Mobile phone inspector utility generates complete report of mobile and SIM card phonebook entries, SMS capacity status and all other general information. Cell phone forensic tool displays detailed information which includes mobile manufacture name, mobile model number, mobile IMEI number, SIM IMSI number, signal quality and battery status of any supported mobile phone. Mobile phone investigation program supports all major bands of windows operating system based mobile phones also supports SIM card of any connection provider like Airtel, Vodafone, Idea etc. • Platform: Windows • Publisher: • Date: • Size: 716 KB. Mobile phone inspector software is available with Microsoft VC++ source code and can be easily extract all important information related to cell phone and SIM card including mobile manufacturer name, model number, mobile IMEI number, SIM IMSI number, battery status, signal quality etc. PDA investigation tool examines text messages, phonebook numbers, Mobile memory status, SMS card memory status from all major brands of mobile handsets such as Nokia, Samsung, Sony, LG, Motorola, Apple, iMate etc. Mobile phone forensic software is useful for all of those users and developers those are working on ActiveSync, MFC, VC++, embedded VC++, C++ builders and some other development platforms. • Platform: Windows • Publisher: • Date: • Size: 716 KB. Cell phone forensic software is freeware utility that easily extracts your entire mobile and sim related data including IMEI number, sim IMSI number, phonebook entries with name and number, text message of all Symbian OS based Nokia mobile phones and other supported mobile devices. Mobile phone investigation application with source code in Microsoft Visual C++, MFC, embedded C++ is useful for organizations working on AT+CPBR, AT+CBS, AT+CSQ, AT+CIMI and many mobile technologies. ![]() Smart phone inspection program is useful for developers to take detail knowledge about various functions related to mobile phones such as CeCreateFile, CeCreateProcess, CeReadFile, CeGetDeviceId, CeFindAllFiles, CeRegEnumKeyEx and CeRegOpenKey etc. • Platform: Misc Scripts • Publisher: • Date. Phone Spy Software is a stealth spy software which allows you to secretly record all activities of your smart phone. Phone Spy Software alows you to listen to actual phone calls and record every SMS and log every call. To perform initial activation/setup on your. Home > Support > LG > LG Revere 3 > Perform Over-The-Air (OTA) Activation. Verizon LTE Internet and Home Phone. If you've recently purchased a new Verizon cell phone, you must activate or program the. How to Program a Verizon Cell Phone or Update. Cellular phone provider. Access Wireless is a government-supported program that ensures anyone can get a mobile phone and free or affordable cell phone service. ![]() ![]() All the calls and SMS logs are uploaded to your online account. Phone Spy Software starts automatically at every phone boot and records every SMS message sent or received, logs all inbound and outbound phone calls, records the visited websites and many more. Basically, all the actions performed on the phone are recorded. • Platform: Android, BlackBerry, Handheld, iOS 4.x, Pocket PC, Symbian, Windows • Publisher: • Date: • Size: 78 KB. Cell Phone Spy - ClubMz, a leading spy company from USA is offering a huge discount this Christmas on its best selling cell phone spy software called e-spy. A software that allows you to spy on any other cell phone allowing you to Listen/Record calls and short messages (SMS). The program is called 'Extreme e-Spy' and will cost you only $ 15.00 USD till Christmas, as the offer is for limited time only and normaly the software costs is much Higher. ClubMz, the manufacturers usually praise the product as a means to catch cheating partner or to monitor your teenagers or employees but one cannot deny its bad usage Once bought you'll need to. • Platform: Java, Mac, OS/2, Palm, Pocket PC, RIM, Symbian, Windows • Publisher: • Date: • Size: 5130 KB. Simple Phone Book was designed to be a simple, easy to use phone book software Simple Phone Book was designed to be a simple, easy to use phone book software. All it does is store data. It's not the worlds most advanced and techie address book. It's a simple way to keep track of numbers and whatever else you want. Here are some key features of 'Simple Phone Book':Add unlimited entries and custom fields.Custom colors for each individual type of entry for easy reading (e-mail can be blue, home phone can be yellow, etc. • Platform: WinOther • Publisher: • Date: • Size: 114 KB. ![]() Spy SMS software is the next generation of smart phone spy software. It allows you to monitor SMS activities of a particular target cell phone. If installed on the target mobile phone, it will automatically copy all text messages (SMS) sent and received by the target mobile phone and silently forward these copied messages to your own mobile phone in real-time, all the time without the target user knowing it. The Spy SMS software also allows you to receive copies of SMS messages sent and received by your target cell phone during certain times of the day or for a short or accurate periods, as you can activate or deactivate the SMS forwarding feature at any time you wish. • Platform: Symbian • Publisher: • Date. WirelessTone(GPRS Surfing Card Software)can provide multiple functions for your wireless surfing card without. Wireless Tone( wireless surfing card software) can provide multiple functions for your wireless surfing card without being restricted to network dialing platform. As extraordinary wireless surfing card software, Wireless Tone supports all the GPRS wireless surfing card; Wireless Tone does not occupy any of your memory or CPU; it can fulfill such tasks as equipment management, MMS editing, number dialing, contact person management, and sending and receiving MMS and short messages. • Platform: Windows • Publisher: • Date: • Size: 9103 KB. Pimp-up your Phone! Free software to enhance your phone, songs, ringtones and wallpapers. Get the latest sounds and stuff for the latest phones. We bring you the largest collection of polyphonic ringtones and monophonic ringtones, graphics, games and software. All ringtones and graphics have previews. Huge range of polyphonic, real music and video ringtones as well as monophonic and free composer ringtones for Nokia, all the latest hits as realtones, monophonic & polyphonic ringtones! • Platform: Windows • Publisher: • Date: • Size: 1008 KB. The GEOTEK Phone Book software is the most popular and advanced Asterisk Phone Book on the market The GEOTEK Phone Book software is the most popular and advanced Asterisk Phone Book on the market. In addition to the phone book features, it also contains Web Dialer, Caller History, Voicemail and Mobile applications.It installs in minutes on any Asterisk server and does not interfere with management software, such as PBX Manager, FOP, Trixbox, or other popular management tools. It has a cute, ergonomically designed web interface that allows to look up phone numbers and to modify and update the Asterisk caller database, so that callers can be identified by name on all SIP telephones. • Platform: WinOther • Publisher: • Date: • Size: 2048 KB. Eahoosoft DVD to Mobile Phone Converter is professional DVD to Mobile Phone Converter software, which helps you rip and convert DVD to Mobile Phone 3GP, MP4, AVI, 3G2 and other video and audio formats. The output videos/audios can be played on most popular Mobile Phones including Nokia, Samsung, Motorola, LG, Sony Ericsson, Palm, etc. Compared with other DVD to Mobile Phone ripping software, Eahoosoft DVD to Mobile Phone Converter possesses more powerful editing functions. When converting DVD to Mobile Phone, you can adjust output video effect, trim any part of DVD, crop the video play region and specify the output settings, select target subtitle and audio track and so on. • Platform: Windows • Publisher: • Date: • Size: 7486 KB. Daniusoft DVD to Mobile Phone Converter is a powerful and easy-to-use DVD to Mobile Phone converter software which can convert DVD to Mobile Phone video including 3GP, MP4, AVI, WMV which fits for Palm, LG, Nokia, Motorola, Samsung, Pocket PC, Sony Erission and other popular mobile players. This great DVD to Mobile Phone Converter can convert DVD to Mobile Phone Video in a super fast Converter speed and has lots of powerful functions, so that you can enjoy your DVDs on your Mobile Phone anytime and anywhere! This best Mobile Phone Converter provides you multifunctional conversion, convert DVD to Mobile Phone by custom file size, rip DVD's any segment, select target subtitle and audio track and trim or crop DVD with only a few clicks. • Platform: Pocket PC, Windows • Publisher: • Date: • Size: 6184 KB. DDVideo DVD to Cell Phone Converter Gain,a powerful DVD to Cell Phone Ripper software,which can rip and convert DVD movies to Cell Phone MP4,H.264/MPEG-4(.mp4),AVI,WMV,3GP video format and AAC, MP3,M4A,MP2,WAV, WMA audio format for all cell phone like LG, Nokia, Motorola, Samsung, Pocket PC, Sony XPeria, BlackBerry, Google Phone etc. DDVideo DVD to Cell Phone Converter Gain is also designed as the video editor for you to recreate the video as you like to enrich your enjoyment of any cell phone, including: *Gain TargetVolume you want:set a output audio gain decibel number for specified dvd video without loss of original sound quality recommend is 89 db(Friendly Reminder:The sound too large,our hearing will be hurt,please care for health care hearing. • Platform: Windows • Publisher: • Date: • Size: 10803 KB. How to get complete saved information of your mobile phone within few minutes? PDA investigation program provides all information of your mobile phone in just single click. Free download cell phone inspector can easily be installed on Windows Operating System including windows XP, Vista, 2003, 2000, NT, ME 98. Mobile phone forensic application allows you to gather all significant information about your cell phone including phone book details with contact name number, SMS information with sender number, date time details. • Platform: Windows • Publisher: • Date: • Size: 716 KB. Watch video's and movies on your phone. This software lets you convert movies and video's to your smartphone and watch it in great quality in full screen landscape mode. A memorycard as small as 128 Mb is sufficient to store a full length feature film. You can watch your content on the go, on any smartphone, from a postage stamp size memory card. You can take any video (AVI, MPEG, DV) from any drive (harddisk, CD, DVD or removable and put it on any Windows Mobile smartphone. Take blockbuster movies with you while you travel, or let the kids watch their favourite cartoons in the car. • Platform: Windows • Publisher: • Date: • Size: 2929 KB. Call recorder, Answering machine, Caller ID, Dialer, and Phone book software. Record outgoing or incoming call or any signal on the phone line. Auto answer machine mode will anwer the incoming calls and play optional greeting legal message before recording. Photo caller ID will display the caller name, number and the photo from the phone book. Lots of nice features and easy user interface.Hundreds of hours of telephone conversations can be stored on your hard drive. All recorded phone calls are compressed to maximize storage capacity. • Platform: Windows • Publisher: • Date: • Size: 8990 KB. Cell phone investigation utility capable to gather complete information from mobile like SMS, contact details, mobile IMEI number, sim IMSI number, mobile model number, mobile manufacturer name, signal quality and battery status etc. Mobile phone examiner with source code in Visual C++, MFC, ActiveSync and DLL also fetches details from all Windows mobile phones, Pocket PC, PDA and Smart phone devices.Application analyzes mobile hardware device connected to USB port of computer. Cell phone inspection software helps in studying different mobile functions like CeCreateProcess, CeCreateFile, CeReadFile, CeGetDeviceId, CeRapiInit, CeGetSystemInfo, CeGetStoreInformation, CeGetVersionEx, CeFindAllFiles, CeRegEnumKeyEx, CeRegOpenKeyEx, CeFindAllDatabase and many more. • Platform: Windows • Publisher: • Date: 18-9-2009 • Size: 720 KB. The RT- Phone Employee Phone Directory Software is a browser-based solution that makes keeping an online phone directory quick and easy. With the RT- Phone Employee Phone Directory Software you can provide employees with current contact information, eliminate the hassle of keeping printed phone books, and easily locate employee phone numbers and office info. Supported web server operating systems include Microsoft IIS. This version is good for up to 100 directory entries. Games Yard - Best Online Games Collection Site. • Platform: • Publisher: • Date. RecOnPhone - Full-featured phone recording software with Caller Id, Email, DTMF, Editing RecOnPhone is a phone recording application that uses a modem or a RecOnBox.While in automatic mode, the software detects each incoming or out going phone call, it automatically records it and stops when the call is over. Audio recordings are compressed and stored in the hard drive. Listening is done with navigation buttons, you change from one record to another and also navigate within a record, going 5 or 30 seconds forward or backward. • Platform: WinOther • Publisher: • Date: 27-6-2009 • Size: 1423 KB. New Reviews It is quite difficult to find any computer users nowadays who do not need to use the web, for any purpose. Whether it is for education or entertainment, the web is your best resource. You also need to use various online services for banking. Nowadays, a majority of PC users resort to digital image editing once in a while. It may be required for retouching image captured in smart phones or for creating an album in Facebook. For some people, tinkering with digital images is a pastime too. Nearly every PC users need to take screenshots from time to time, whether it is for personal or professional needs. While using the basic Windows screenshot capture method is available, it is not adequate for everyone. When you want to capture. 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No serious PC user needs introduction about the need to backup data, both at home and workplace. There was a time when majority of people used external HDD or storage media to backup important data. However, local data backup has its limitations and. To keep your laptop and desktop running smoothly and get the best performance out of it, it is important to pay attention to some aspects. It is quite important to ensure your PC device drivers are updated. Components like motherboards, GPU and. New Downloads True Burner is a compact, easy to use and powerful burning software, which enables you to make and burn standard. Advanced SystemCare Ultimate 11 is a powerful and full-scale software for PC security and performance. Radmin VPN - is a free and easy-to-use software product to create virtual private network (VPN). The program allows users. Huella Dactilar SDK allows you to integrate fingerprint recognition technology in various types of applications with just. Specially designed for small to medium work groups to share data! To casting your unique corporate culture! Universal IPv4/IPv6 scanner. Can ping computers, scan ports, discover shared folders and retrieve practically any. Jar2Exe is a tool to wrap jar files into binary files for Windows, Linux and Mac. It supports 32 bits and 64 bits. Free convert PDF to Word, DOCX, RTF, HTML, Excel Transform PDF documents to image formats like JPEG, PNG, TIFF, BMP, and. Video quality test software for QA, technical support, and system integration teams offering key features of StreamEye. SecureCRT is a rock-solid terminal emulator with the strong encryption and authentication options of the Secure Shell. Hi, I'm trying to activate a refurbished Samsung Galaxy S2 with my newly ported number, but my phone keeps thinking it's roaming. I've brought my phone to several different locations (all in Washington DC, where I live) and it consistently thinks I'm roaming. I've had tech support walk me through the manual reboot, and nothing seems to work. When I type in the MLS code, it says 'Invalid Lock Code.' I had a second Samsung Galaxy S2 shipped to me, but it is doing the same thing. I think I stumped both of the support reps I spoke with. The guy I spoke with today said it might be because I ported a Seattle area code (where we used to live). Anyone had experience with this? It couldn't possibly be that Sprint does not have data coverage where I am. I'm fine with getting a new number.
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We describe a bacterial two-hybrid system that allows an. Cyclic AMP then triggers transcriptional activation of. Design of a Two-Hybrid System Based on. A Bacterial Two-Hybrid System Based on Transcription Activation A Bacterial Two-Hybrid System Based on Transcription Activation Dove, Simon L.; Hochschild, Ann 2004-01-01 00:00:00 We describe the use of a bacterial two-hybrid system for the study of protein-protein interactions in Escherichia coli. The bacterial two‐hybrid (BACTH, for “Bacterial Adenylate Cyclase‐based Two‐Hybrid”) technique is a simple and fast genetic approach to analyze protein‐protein interactions in vivo. In this system, the proteins of interest are genetically fused to two complementary fragments from the catalytic domain of Bordetella pertussis adenylate cyclase and co‐expressed in strains of Escherichia coli deficient in adenylate cyclase. ![]() ![]() Association of the hybrid proteins restores synthesis of cyclic AMP (cAMP), which then triggers the expression of catabolic operons such as the lactose operon or the maltose regulon. As BACTH uses a cAMP second messenger, the association between the chimeric proteins can take place at a distance from the transcription machinery. This technique is therefore particularly appropriate for studying interactions involving integral‐membrane or membrane‐associated proteins that may not be soluble in the cytoplasm, and/or that may only associate in the plane of the membrane. ![]() This unit describes the basic procedures to characterize protein‐protein interactions with the BACTH genetic system and to search for potential partners of known proteins. © 2017 by John Wiley & Sons, Inc. Keywords: protein interaction assay; two‐hybrid technique; membrane protein; library screening; Escherichia coli; cAMP signaling. D., & Grossmann, B. Digestion of DNA with restriction endonucleases. ![]() Current Protocols in Molecular Biology, 31, 3.1.1–3.1.21. Busby, S., & Ebright, R. Transcription activation by catabolite activator protein (CAP). Journal of Molecular Biology, 293, 199–213. Doi: 10.1006/jmbi.1999.3161 Chen, C., Banga, S., Mertens, K., Weber, M. ![]() M., Gorbaslieva, I., Tan, Y., Samuel, J. 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GraXSR proteins interact with the VraFG ABC transporter to form a five‐component system required for cationic antimicrobial peptide sensing and resistance in Staphylococcus aureus. Antimicrobial Agents and Chemotherapy, 56, 1047–1058. Doi: 10.1128/AAC.05054‐11 Fields, S., & Song, O. A novel genetic system to detect protein‐protein interactions. Nature, 340, 245–246. Doi: 10.1038/340245a0 Finley, R. J., & Brent, R. Interaction mating reveals binary and ternary connections between Drosophila cell cycle regulators. Proceedings of the National Academy of Sciences of the United States of America, 91, 4. Doi: 10.1073/pnas.0 Fransen, M., Brees, C., Ghys, K., Amery, L., Mannaerts, G. P., Ladant, D., & Van Veldhoven, P. Analysis of mammalian peroxin interactions using a non‐transcription‐based bacterial two‐hybrid assay. Molecular & Cellular Proteomics, 1, 243–252. Doi: 10.1074/mcp.M100025‐MCP200 Gauliard, E., Ouellette, S. P., Rueden, K. J., & Ladant, D. 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The beta‐lactam resistance protein Blr, a small membrane polypeptide, is a component of the Escherichia coli cell division machinery. Journal of Bacteriology, 194, 5576–5588. Doi: 10.1128/JB.00774‐12 Karimova, G., Pidoux, J., Ullmann, A., & Ladant, D. A bacterial two‐hybrid system based on a reconstituted signal transduction pathway. Proceedings of the National Academy of Sciences of the United States of America, 95, 5752–5756. Doi: 10.1073/pnas. Karimova, G., Robichon, C., & Ladant, D. Characterization of YmgF, a 72‐residue inner membrane protein that associates with the Escherichia coli cell division machinery. Journal of Bacteriology, 191, 333–346. Doi: 10.1128/JB.00331‐08 Karimova, G., Ullmann, A., & Ladant, D. A bacterial two‐hybrid system that exploits a cAMP signaling cascade in Escherichia coli. Methods in Enzymology, 328, 59–73. Doi: 10.1016/S0076‐680‐0 Karimova, G., Ullmann, A., & Ladant, D. Protein‐protein interaction between Bacillus stearothermophilus tyrosyl‐tRNA synthetase subdomains revealed by a bacterial two‐hybrid system. Journal of Molecular Microbiology and Biotechnology, 3, 73–82. Kleinschnitz, E.‐M., Heichlinger, A., Schirner, K., Winkler, J., Latus, A., Maldener, I., Muth, G. Proteins encoded by the mre gene cluster in Streptomyces coelicolor A3(2) cooperate in spore wall synthesis. Molecular Microbiology, 79, 1367–1379. Doi: 10.1111/j.1365‐2958.2010.07529.x Kramer, M. F., & Coen, D. Enzymatic amplification of DNA by PCR: Standard procedures and optimization. Current Protocols in Molecular Biology, 56, 15.1.1–15.1.14. B., & Nichols, N. DNA‐dependent DNA polymerases. Current Protocols in Molecular Biology, 84, 3.5.1–3.5.19. Ladant, D., & Ullmann, A. Bordetella pertussis adenylate cyclase: A toxin with multiple talents. Trends in Microbiology, 7, 172–176. Doi: 10.1016/S0966‐842X(99)01468‐7 Miller, J. A short course in bacterial genetics: A laboratory manual and handbook for Escherichia coli and related bacteria. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press. E., Oshima, T., Piper, S. E., Webster, C. L., Westblade, L. F., Karimova, G., Lee, D. The Escherichia coli regulator of sigma 70 protein, Rsd, can up‐regulate some stress‐dependent promoters by sequestering sigma 70. Journal of Bacteriology, 189, 3489–3495. Doi: 10.1128/JB.00019‐07 Nichols, N. Current Protocols in Molecular Biology, 93, 3.12.1–3.12.7. Ouellette, S. P., Gauliard, E., Antosova, Z., & Ladant, D. A Gateway‐compatible bacterial adenylate cyclase‐based two‐hybrid system. Environmental Microbiology Reports, 6, 259–267. Doi: 10.1111/1758‐2229.12123 Ouellette, S. P., Karimova, G., Subtil, A., & Ladant, D. Chlamydia co‐opts the rod shape‐determining proteins MreB and Pbp2 for cell division. Molecular Microbiology, 85, 164–178. Doi: 10.1111/j.1365‐2958.2012.08100.x Ouellette, S. P., Rueden, K. J., Gauliard, E., Persons, L., de Boer, P. A., & Ladant, D. Analysis of MreB interactors in Chlamydia reveals a RodZ homolog but fails to detect an interaction with MraY. Frontiers in Microbiology, 5, 279. Doi: 10.3389/fmicb.2014.00279 Paschos, A., den Hartigh, A., Smith, M. A., Atluri, V. L., Sivanesan, D., Tsolis, R. M., & Baron, C. An in vivo high‐throughput screening approach targeting the type IV secretion system component VirB8 identified inhibitors of Brucella abortus 2308 proliferation. Infection and Immunity, 79, 1033–1043. Doi: 10.1128/IAI.00993‐10 Pfeiffer, D., & Jendrossek, D. Interaction between poly(3‐hydroxybutyrate) granule‐associated proteins as revealed by two‐hybrid analysis and identification of a new phasin in Ralstonia eutropha H16. Microbiology (Reading, England), 157, 2795–2807. Doi: 10.1099/mic.0.051508‐0 Robichon, C., Karimova, G., Beckwith, J., & Ladant, D. Role of leucine zipper motifs in association of the Escherichia coli cell division proteins FtsL and FtsB. Journal of Bacteriology, 193, 4988–4992. Doi: 10.1128/JB.00324‐11 Sambrook, J., Russell, D. W., & Sambrook, J. The condensed protocols from molecular cloning: A laboratory manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press. Sands, B., & Brent, R. Overview of post Cohen‐Boyer methods for single segment cloning and for multisegment DNA assembly. Current Protocols in Molecular Biology, 113, 3.26.1–3.26.20. Doi: 10.10727.mb0326s113 Seidman, C. E., Struhl, K., Sheen, J., & Jessen, T. Introduction of plasmid DNA into cells. Current Protocols in Molecular Biology, 37, 1.8.1–1.8.10. Stynen, B., Tournu, H., Tavernier, J., & Van Dijck, P. Diversity in genetic in vivo methods for protein‐protein interaction studies: From the yeast two‐hybrid system to the mammalian split‐luciferase system. Microbiology and Molecular Biology Reviews, 76, 331–382. Doi: 10.1128/MMBR.05021‐11 Voytas, D. Agarose gel electrophoresis. Current Protocols in Molecular Biology, 51, 2.5A.1–2.5A.9. Preparation of genomic DNA from bacteria. Current Protocols in Molecular Biology, 00, 2.4.1–2.4.5. S., Gyuris, J., & Brent, R. Mxi1, a protein that specifically interacts with Max to bind Myc‐Max recognition sites. Cell, 72, 223–232. Doi: 10.1016/0092‐862‐A Zoued, A., Durand, E., Brunet, Y. R., Spinelli, S., Douzi, B., Guzzo, M., Cascales, E. Priming and polymerization of a bacterial contractile tail structure. Nature, 531, 59–63. Doi: 10.1038/nature17182 Internet Resources Euromedex: Commercial supplier of the BACTH system. Supplier for Gateway ‘recombineering’ cloning technique. ABSTRACT A number of transcriptional regulators mediate their effects through direct contact with the ς 70 subunit of Escherichia coli RNA polymerase (RNAP). In particular, several regulators have been shown to contact a C-terminal portion of ς 70 that harbors conserved region 4. This region of ς contains a putative helix-turn-helix DNA-binding motif that contacts the −35 element of ς 70-dependent promoters directly. Here we report the use of a recently developed bacterial two-hybrid system to study the interaction between the putative anti-ς factor Rsd and the ς 70 subunit of E. Using this system, we found that Rsd can interact with an 86-amino-acid C-terminal fragment of ς 70 and also that amino acid substitution R596H, within region 4 of ς 70, weakens this interaction. We demonstrated the specificity of this effect by showing that substitution R596H does not weaken the interaction between ς and two other regulators shown previously to contact region 4 of ς 70. We also demonstrated that AlgQ, a homolog of Rsd that positively regulates virulence gene expression in Pseudomonas aeruginosa, can contact the C-terminal region of the ς 70 subunit of RNAP from this organism. We found that amino acid substitution R600H in ς 70 from P. Aeruginosa, corresponding to the R596H substitution in E. Coli ς 70, specifically weakens the interaction between AlgQ and ς 70. Taken together, our findings suggest that Rsd and AlgQ contact similar surfaces of RNAP present in region 4 of ς 70 and probably regulate gene expression through this contact. Sigma factors are subunits of bacterial RNA polymerase (RNAP) that direct the holoenzymes that contain them to promoters of a specific class (). In Escherichia coli there are seven different species of ς factors, and ς 70 is the principal ς factor (). The presence of different types of ς factors within a cell with distinct DNA sequence binding specificities provides a mechanism for coordinate regulation of genes that are controlled by promoters of the same class. Competition between different ς factors for the available RNAP core enzyme in part determines which genes are transcribed within a cell at any given time (). This competition can be influenced by anti-ς factors, which are regulatory proteins that bind ς factors and often prevent their association with the RNAP core enzyme (, ). Anti-ς factors ultimately inhibit transcription from the class of promoters recognized by the ς factors that they sequester. The ς 70 subunit of RNAP participates in a number of protein-protein interactions, including interactions with other subunits of the polymerase complex (, ) and interactions with transcriptional regulators (, ). The regulators that interact with ς 70 often contact a region of ς that contains a putative helix-turn-helix DNA-binding motif responsible for contacting the −35 elements of ς 70-dependent promoters (,,). This DNA-binding region of ς is conserved in members of the ς 70 family of proteins and is called region 4 (). Recently, Jishage and Ishihama identified a protein in E. Coli that was preferentially made by cells during the stationary phase of growth and was associated with the ς 70subunit of RNAP in stationary-phase extracts (). The protein was named Rsd (which stands for regulator of sigma D) since it was found to associate specifically with ς 70(but not with several alternative ς factors) and was shown to be capable of inhibiting ς 70-dependent transcription from certain promoters in vitro (). The binding site for Rsd on ς 70 was mapped to a C-terminal tryptic fragment encompassing conserved region 4 (). On the basis of these observations and because the synthesis of Rsd coincides with the general shutdown in ς 70-dependent transcription that occurs as cells enter the stationary phase of growth, Jishage and Ishihama suggested that Rsd might be an anti-ς factor (). Subsequent work has shown that consistent with this idea, Rsd may facilitate the replacement of ς 70 by the stationary-phase-specific ς factor ς 38 in functional RNAP holoenzyme complexes as cells go from the exponential phase to the stationary phase of growth (). The sequence of putative anti-ς factor Rsd is similar to the sequence of a regulator of alginate production in Pseudomonas aeruginosa called AlgQ (or AlgR2) (). Alginate is an important virulence factor that imparts the characteristic mucoid phenotype to P. Aeruginosa isolated from the lungs of cystic fibrosis patients (). Aeruginosa isolates from other sources are typically nonmucoid and do not exhibit activated expression of genes involved in alginate production (). However, the production of alginate is believed to promote survival of P. Aeruginosa in the special environment of the lungs of cystic fibrosis patients, contributing to resistance to both immune responses and antibiotics (). AlgQ was originally identified as a positive transcriptional regulator of the key alginate biosynthetic gene algD (, ), which is expressed at high levels in mucoid cells. The regulation of the algD gene is complex and involves two different ς factors; transcription initiates from two superimposed promoters, one of which is recognized by RNAP containing ς E (AlgU/AlgT) and the other of which is recognized by RNAP containing ς 54 (RpoN) (,,,,,,). Furthermore, at least one DNA-binding protein, AlgR (AlgR1), is known to bind to specific sites upstream of the algD promoter and activate transcription (,,). The mechanism by which AlgQ positively regulates transcription of the algD gene is not known. We were interested in testing the idea that like Rsd, AlgQ interacts with region 4 of the ς 70 subunit of RNAP. In this study we tested this idea explicitly by using a bacterial two-hybrid system. We found that both Rsd and AlgQ can interact with a ς 70 moiety encompassing region 4 in vivo, and we identified an amino acid substitution in region 4 that specifically weakens the interaction of Rsd and AlgQ with the ς moiety. Plasmids and strains. Coli XL1-blue (Stratagene) was used as the recipient strain for all plasmid constructions. Coli KS1 harbors on its chromosome the lac promoter derivative p lacO R2-62 driving expression of a linked lacZ reporter gene and has been described previously (). Coli SF1 harbors an F′ episome containing the lac promoter derivative p lacO R2-55/Cons −35 driving expression of a linked lacZ reporter gene and has also been described previously (). Plasmids pACλcI, pACΔcI, pBRα, pBRα-ς 38, pBRα-ς 70, and pBRα-ς 70 (R596H) have all been described previously (, ). Plasmid pACλcI-Rsd encodes λcI (residues 1 to 236) fused to Rsd (residues 1 to 158) via three alanine residues. PACλcI-Rsd was made by cloning the appropriate NotI- BamHI-digested PCR product into NotI- BstYI-digested pACλcI32 (); expression of the cI-rsd fusion gene was therefore under the control of the lacUV 5 promoter. Plasmid pACλcI-AlgQ encodes λcI (residues 1 to 236) fused to AlgQ (residues 1 to 160) via three alanine residues, and it was made in a manner similar to the manner in which pACλcI-Rsd was made. Expression of the cI-algQ fusion gene on pACλcI-AlgQ is also under the control of the lacUV5 promoter. Plasmid pACλcI-AsiA encodes λcI (residues 1 to 236) fused to AsiA from bacteriophage T4 (residues 1 to 90) via three alanine residues, and it was made in a manner similar to the manner in which pACλcI-Rsd was made. Plasmid pACΔ-35λcI-AsiA contains the cI-asiA fusion gene from plasmid pACλcI-AsiA under the control of a lacUV5 promoter variant in which the −35 element of the promoter has been deleted. Plasmid pACΔ-35λcI-AsiA, therefore, expresses less of the λcI-AsiA fusion protein than plasmid pACλcI-AsiA expresses under identical conditions. PACΔ-35λcI-AsiA was made by cloning the appropriate HindIII- BstYI fragment from pACλcI-AsiA into plasmid pA3B2 () cut with both HindIII and BstYI. Plasmid pBRα-ς 70PA encodes residues 1 to 248 of the α subunit of E. Coli RNAP fused to residues 532 to 617 of the ς 70 subunit of P. The hybrid α-ς 70PAgene was made by performing PCR and was cloned into HindIII- BamHI-digested pBRα. Expression of the chimeric gene on pBRα-ς 70PA is, therefore, under the control of tandem lpp and lacUV5 promoters. PBRα-ς 70PA (R600H) is a derivative of pBRα-ς 70PA in which the R600H substitution in the ς moiety of the chimera was introduced by PCR. The lac promoter derivative p lacCOP-93+O L2-62 was made by the PCR and contains two λ operators; a near-consensus λ operator (TACCACCGGCGGTGATA) and O L2 (CAACACCGCCAGAGATA) are centered 93 and 62 bp, respectively, upstream of the transcriptional start site of the lac core promoter. Plasmid pFW11-COP-93+O L2-62 was constructed by cloning an EcoRI- HindIII-cut PCR product containing p lacCOP-93+O L2-62 into pFW11 () cut with EcoRI and HindIII. Plasmid pFW11-COP-93+O L2-62 was then transformed into strain CSH100, and the promoter- lacZ fusion was recombined onto an F′ episome and mated into strain FW102 () to create reporter strain F′93+62. The PCR-amplified regions of all plasmids were sequenced to confirm that no errors had been introduced as a result of the PCR process. Experimental procedures.Cells were grown in LB supplemented with kanamycin (50 μg/ml), chloramphenicol (25 μg/ml), carbenicillin (50 μg/ml), and isopropyl-β- d-thiogalactoside (IPTG) at the concentration indicated. Cells were permeabilized with sodium dodecyl sulfate-CHCl 3 and assayed for β-galactosidase activity essentially as described previously (). Assays were performed at least three times in duplicate on separate occasions, and representative data sets are shown below. The values are averages based on one experiment; duplicate measurements differed by less than 10%. ΛcI-Rsd activates transcription from a test promoter in the presence of a chimeric α-subunit harboring region 4 of E. Coli ς 70.We sought to detect an interaction between Rsd and region 4 of ς 70 in vivo by using a bacterial two-hybrid system that we had recently developed (, ). This bacterial two-hybrid system is based on the finding that any sufficiently strong interaction between a protein bound upstream of a suitable test promoter and a component of RNAP can activate transcription in E. Thus, two proteins that interact with one another can mediate transcriptional activation in E. Coli provided that one protein is fused to a DNA-binding protein and the other is fused to a component of RNAP (, ). Our strategy for detecting an interaction between Rsd and region 4 of ς 70 involved the use of two chimeric proteins, one comprising Rsd fused to the repressor of bacteriophage λ (λcI) and the other comprising a modified form of the α-subunit of RNAP in which the C-terminal domain (CTD) of α has been replaced by a C-terminal fragment of ς 70. We reasoned that RNAP containing the resulting α-ς 70 chimera would display a target for Rsd that could be contacted by a DNA-bound λcI-Rsd dimer (Fig. Having fused the entire Rsd protein (residues 1 to 158) to the C terminus of λcI, we placed the gene encoding this chimeric protein on a plasmid vector downstream of the IPTG-inducible lacUV5 promoter, thus creating plasmid pACλcI-Rsd. We used plasmid pBRα-ς 70 as a source of the α-ς 70 chimera. This plasmid encodes a chimera in which residues 528 to 613 of E. Coliς 70 are fused to residues 1 to 248 of the α-subunit of RNAP (). We introduced plasmids pACλcI-Rsd and pBRα-ς 70 into E. Coli KS1 (), which harbors on its chromosome the lac promoter derivative p lacO R2-62 (bearing a single λ operator centered 62 bp upstream of the transcriptional start site) linked to a lacZ reporter gene. We then tested the ability of the λcI-Rsd chimera to activate transcription from the p lacO R2-62 test promoter in the presence of the α-ς 70 chimera. Figure B shows that λcI-Rsd activated transcription from the test promoter up to ∼24-fold in cells containing the α-ς 70 chimera compared to control cells containing only wild-type α. An additional control revealed that λcI (lacking the fused Rsd moiety) did not activate transcription from the test promoter in the presence of the α-ς 70 chimera (Fig. We also found that λcI-Rsd did not activate transcription from the test promoter in the presence of an α-ς 38 chimera (comprising residues 1 to 248 of α fused to residues 243 to 330 of ς 38) encoded by plasmid pBRα-ς 38 (; data not shown). Transcriptional activation by λcI-Rsd in the presence of the α-ς 70 chimera. (A) Replacement of the RNAP α-CTD by a C-terminal fragment of E. Coliς 70 (residues 528 to 613) permits interaction with the Rsd moiety of a λcI-Rsd chimera bound to DNA. The diagram depicts the test promoter p lacO R2-62, which bears the λ operator O R2 centered 62 bp upstream from the transcriptional start site of the lac core promoter. In reporter strain KS1 the p lacO R2-62 test promoter is located on the chromosome and drives expression of a linked lacZ gene. The N-terminal domain of α is designated αNTD. (B) Effect of λcI-Rsd on transcription in vivo from p lacO R2-62 in the presence of the α-ς 70 or α-ς 70(R596H) chimera. KS1 cells harboring compatible plasmids expressing the indicated proteins were grown in the presence of different concentrations of IPTG and assayed for β-galactosidase activity. Substitution R596H in the ς moiety of the α-ς 70chimera weakens the interaction between ς and RsdOur ability to link the protein-protein interaction between Rsd and its target on ς 70 to transcriptional activation provided us with a useful genetic tool for dissecting this interaction. We were particularly interested in identifying mutant forms of ς 70 that were specifically defective in the ability to interact with Rsd. We therefore introduced a variety of amino acid substitutions into the ς moiety of the α-ς 70 chimera and tested the effects of these substitutions on the ability of the λcI-Rsd fusion protein to mediate transcriptional activation from the test promoter. Figure B shows that substitution R596H in the ς moiety of the α-ς 70chimera strongly reduced the magnitude of λcI-Rsd-dependent activation (to a factor of ∼5). Substitution R596A in the ς moiety of the α-ς 70 chimera had a nearly identical effect on the magnitude of the activation (data not shown). In contrast, substitutions L573A, E591A, E591Q, H600A, and H600R did not decrease the magnitude of λcI-Rsd-dependent activation (data not shown). Substitution R596H in the ς moiety of the α-ς 70chimera does not compromise the ability of a superactivating variant of λcI to stimulate transcription from an appropriate test promoter.To determine whether the R596H substitution affects the interaction of ς 70 with Rsd specifically, we tested its effect on the ability of another protein to interact with the ς moiety of the chimera. We showed recently that the λcI protein (a transcriptional activator as well as a repressor) can interact specifically with the ς moiety of the α-ς 70 chimera and stabilize its binding to a promoter −35 element (). The in vivo assay which we designed to detect the interaction between λcI and region 4 of ς 70 is shown in Fig.A. In this experimental setup a DNA-bound λcI dimer activates transcription from test promoter p lacO R2-55/Cons-35 by stabilizing the binding of the ς moiety of the α-ς 70chimera to the ectopic −35 element present upstream of the core promoter elements () (Fig. Transcriptional activation from this test promoter is dependent not only on the protein-protein interaction between λcI and the tethered ς moiety but also on the protein-DNA interaction between the tethered ς moiety and the ectopic −35 element (). Transcriptional activation by λcI superactivator in the presence of the α-ς 70 chimera. (A) λcISa109 stabilizes the binding of region 4 of ς 70 to an ectopic −35 element. The diagram depicts the test promoter p lacO R2-55/Cons-35, which bears an ectopic −35 element and the λ operator O R2 centered 45.5 and 55 bp, respectively, upstream from the transcriptional start site of the lac core promoter. In reporter strain SF1 the p lacO R2-55/Cons-35 test promoter is located on an F′ episome and drives expression of a linked lacZgene. (B) Effect of substitution R596H in the ς moiety of the α-ς 70 chimera on the ability of λcISa109 to activate transcription in vivo from p lacO R2-55/Cons-35. SF1 cells harboring compatible plasmids expressing the indicated proteins were grown in the presence of different concentrations of IPTG and assayed for β-galactosidase activity. We tested whether the R596H substitution in the α-ς 70 chimera has an effect on the ability of a superactiviating variant of λcI to activate transcription from test promoter p lacO R2-55/Cons-35 (Fig. Reporter strain SF1 carries promoter p lacO R2-55/Cons-35 fused to the lacZ gene in single copy on an F′ episome (). We assayed the ability of λcI superactivator 109 (λcISa109) (, ) to activate transcription from this reporter in the presence of the α-ς 70 chimera with or without the R596H substitution in the ς moiety. ΛcISa109 activated transcription a maximum of ∼5.5-fold from the test promoter in the presence of the α-ς 70 chimera and a maximum of ∼7.5-fold in the presence of the chimera harboring the R596H substitution (Fig. (Although the R596H substitution has previously been shown to inhibit the ability of wild-type λcI to activate transcription from P RM [], we have observed that this substitution does not inhibit the ability of λcISa109 to activate transcription from P RM[see below].) We concluded that substitution R596H in the ς moiety of the α-ς 70 chimera does not result in a general defect in the ability of the ς moiety to interact with other proteins. We also tested the effect of the R596A substitution in the ς moiety of the α-ς 70 chimera on the ability of λcISa109 to activate transcription from the same test promoter. Unlike the R596H substitution, the R596A substitution significantly reduced the magnitude of the activation by λcISa109 (data not shown). Using another assay, however, we were able to determine that this reduction in activation was not due to a general defect caused by the R596A substitution (see below). ΛcI-AlgQ activates transcription from a test promoter in the presence of a chimeric α-subunit harboring region 4 of P. Aeruginosa ς 70.Jishage and Ishihama () noted that Rsd exhibits 31% identity with the alginate regulatory protein AlgQ (also known as AlgR2) from P. AlgQ was originally identified as a positive regulator of alginate production in P. Aeruginosa (,) but has subsequently been shown to regulate several other gene products in this organism (see below). Very little is known about how AlgQ mediates its effects on gene expression. In order to test the hypothesis that AlgQ, like Rsd, can interact with region 4 of ς 70, we made a cI-algQ fusion gene analogous to the cI-rsdfusion gene. We also made a fusion gene encoding a protein analogous to the α-ς 70 chimera that contained region 4 of ς 70 from P. We fused the entire AlgQ protein (residues 1 to 160) to the C terminus of λcI. We placed the gene encoding this chimeric protein on a plasmid vector downstream of the IPTG-inducible lacUV5 promoter, creating plasmid pACλcI-AlgQ. We then fused residues 532 to 617 of P. Aeruginosa ς 70 (equivalent to residues 528 to 613 of E. Coli ς 70) to residues 1 to 248 of α. The resulting chimera was called α-ς 70PA. The hybrid α-ς 70PA gene encoding this chimera was placed on a plasmid vector downstream of tandemly arranged lpp and lacUV5 promoters, creating plasmid pBRα-ς 70PA. We introduced plasmids pACλcI-AlgQ and pBRα-ς 70PA into E. We then tested the ability of the λcI-AlgQ chimera to activate transcription from p lacO R2-62 in the presence of the α-ς 70PA chimera (Fig.A). Figure B shows that λcI-AlgQ activated transcription from the test promoter a maximum of ∼17-fold in cells containing the α-ς 70PA chimera compared to control cells containing only wild-type α. An additional control revealed that λcI without the fused AlgQ moiety did not activate transcription from the test promoter in the presence of the α-ς 70PA chimera (Fig. Transcriptional activation by λcI-AlgQ in the presence of the α-ς 70PA chimera. (A) Replacement of the RNAP α-CTD by a C-terminal fragment of P. Aeruginosaς 70 (residues 532 to 617) permits interaction with the AlgQ moiety of a λcI-AlgQ chimera bound to DNA. The diagram depicts the test promoter p lacO R2-62, which bears the λ operator O R2 centered 62 bp upstream from the transcriptional start site of the lac core promoter. In reporter strain KS1 the p lacO R2-62 test promoter is located on the chromosome and drives expression of a linked lacZ gene. (B) Effect of λcI-AlgQ on transcription in vivo from p lacO R2-62 in the presence of the α-ς 70PA or α-ς 70PA(R600H) chimera. KS1 cells harboring compatible plasmids expressing the indicated proteins were grown in the presence of different concentrations of IPTG and assayed for β-galactosidase activity. Substitution R600H in the ς moiety of the α-ς 70PA chimera weakens the interaction between ς and AlgQ.The ς 70 subunits from E. Aeruginosa are very similar to one another, exhibiting ∼83% identity over the length of the ς fragments (86 amino acids) that we used in our experiments (). We wanted to test whether substitution R600H in ς 70 from P. Aeruginosa, which corresponds to the R596H substitution in E. Coli ς 70, had any effect on the ability of λcI-AlgQ to activate transcription in the presence of the α-ς 70PA chimera. To do this, we made a version of the α-ς 70PA chimera harboring substitution R600H [α-ς 70PA(R600H)] and assayed the ability of λcI-AlgQ to activate transcription in KS1 cells expressing this chimera. Figure B shows that λcI-AlgQ activated transcription from the reporter gene a maximum of ∼5-fold in the presence of the α-ς 70PA(R600H) chimera, compared to a maximum of ∼17-fold with the chimera derived from the wild-type form of P. Aeruginosa ς 70. Substitution R600H in the ς moiety of the α-ς 70PA chimera does not compromise the ability of a superactivating variant of λcI to stimulate transcription from an appropriate test promoter.In order to assess whether the effect of the R600H substitution was specific for the interaction between ς 70PA and AlgQ, we first tested whether λcISa109 could interact with the ς moiety of the α-ς 70PA chimera. We found that λcISa109 stimulated transcription from test promoter p lacO R2-55/Cons-35 up to ∼3.5-fold specifically in the presence of the α-ς 70PA chimera (Fig. Furthermore, introduction of the R600H substitution into the ς moiety of the α-ς 70PA chimera did not abrogate the stimulatory effect of λcISa109 (instead it resulted in a modest increase in the observed activation), suggesting that the effect of the R600H substitution is specific for the λcI-AlgQ chimera (Fig. Transcriptional activation by λcI superactivator in the presence of the α-ς 70PA chimera. (A) λcISa109 stabilizes the binding of region 4 of ς 70 from P. Aeruginosa to an ectopic −35 element. The diagram depicts the test promoter p lacO R2-55/Cons-35, which bears an ectopic −35 element and the λ operator O R2 centered 45.5 and 55 bp, respectively, upstream from the transcriptional start site of the lac core promoter. In reporter strain SF1 the p lacO R2-55/Cons-35 test promoter is located on an F′ episome and drives expression of a linked lacZ gene. (B) Effect of substitution R600H in the ς moiety of the α-ς 70PA chimera on the ability of λcISa109 to activate transcription in vivo from p lacO R2-55/Cons-35. SF1 cells harboring compatible plasmids expressing the indicated proteins were grown in the presence of different concentrations of IPTG and assayed for β-galactosidase activity. Coli protein Rsd can interact with region 4 of ς 70 from P. Aeruginosa, and the P. Aeruginosa protein AlgQ can interact with region 4 of ς 70 from E. Coli.Given the high degree of similarity between the regions of ς 70from E. Aeruginosa used in our experiments, we thought that each regulator might be able to contact region 4 of ς 70 from either E. We explicitly tested whether Rsd could interact with region 4 of ς 70 from P. Aeruginosa and also whether AlgQ could interact with region 4 of ς 70 from E. FigureA shows that the λcI-Rsd chimera activated transcription from the test promoter in KS1 cells a maximum of ∼52-fold in the presence of the α-ς 70PA chimera, compared to ∼24-fold in the presence of the E. Coli α-ς 70 chimera. We also found that introduction of substitution R600H into the α-ς 70PA chimera reduced the ability of λcI-Rsd to stimulate transcription from the test promoter to a factor of ∼13 (Fig. Rsd can interact with region 4 of ς 70 from P. Aeruginosa, and AlgQ can interact with region 4 of ς 70 from E. (A) Effect of λcI-Rsd on transcription in vivo from p lacO R2-62 in the presence of the α-ς 70PA, α-ς 70PA(R600H), or α-ς 70 chimera. KS1 cells harboring compatible plasmids expressing the indicated proteins were grown in the presence of different concentrations of IPTG and assayed for β-galactosidase activity. (B) Effect of λcI-AlgQ on transcription in vivo from p lacO R2-62 in the presence of the α-ς 70, α-ς 70(R596H), or α-ς 70PA chimera. KS1 cells harboring compatible plasmids expressing the indicated proteins were grown in the presence of different concentrations of IPTG and assayed for β-galactosidase activity. ΛcI-AsiA activates transcription from a test promoter in the presence of either α-ς chimera.We took advantage of another protein known to interact with region 4 of E. Coliς 70 to test further the specificity of the effect of the R596H substitution on the interaction of ς 70 with either Rsd or AlgQ. The AsiA protein encoded by bacteriophage T4 is an anti-ς factor that has been shown to interact specifically and very strongly with region 4 of E. Coli ς 70 (,,,,,). AsiA is thought to work by interacting with ς 70 that is free in solution rather than with ς 70 that is already complexed with the RNAP core enzyme (). The interaction between AsiA and E. Coli ς 70 inhibits the activity of RNAP holoenzyme (containing the AsiA-ς 70complex) by preventing region 4 of ς from contacting the −35 element of ς 70-dependent promoters (,). We therefore sought to detect the interaction between AsiA and region 4 of ς 70 by fusing AsiA to λcI and measuring the ability of the resulting λcI-AsiA chimera to activate transcription from p lacO R2-62 in the presence of either the α-ς 70 chimera or the α-ς 70PA chimera. We fused the entire AsiA protein (residues 1 to 90) to the C terminus of λcI. We placed the gene encoding this chimeric protein on a plasmid vector downstream of the IPTG-inducible lacUV5 promoter, creating plasmid pACλcI-AsiA. Initial experiments demonstrated that the λcI-AsiA chimera was extremely toxic to cells at the levels provided by the expression vector pACλcI-AsiA (data not shown). For subsequent experiments we constructed a different expression vector, pACΔ-35λcI-AsiA, in which the chimeric cI-asiA gene was under control of an IPTG-inducible variant of the lacUV5 promoter that lacked the normal −35 element and therefore directed the synthesis of smaller amounts of the fusion protein. Since pACΔ-35λcI-AsiA made insufficient levels of λcI-AsiA to saturate the λ operator present on the p lacO R2-62 promoter construct, we also constructed a new test promoter bearing two relatively strong λ operators in the upstream region (Fig.A; also see Materials and Methods). Figure B shows that the λcI-AsiA chimera activated transcription from the modified test promoter in the presence of the α-ς 70 chimera derived from ς 70 of either E. Aeruginosa (similar findings with the α-ς 70 chimera from E. Coli have been obtained by S. Hinton [submitted for publication]). The data also show that substitution R596H in the E. Coli ς moiety or the corresponding R600H substitution in the P. Aeruginosa ς moiety had only a modest effect on the ability of the λcI-AsiA chimera to activate transcription from p lacCOP-93+O L2-62. Similarly, the R596A substitution in the E. Coli ς moiety did not reduce the stimulatory effect of the λcI-AsiA chimera (data not shown). Since these substitutions do not appear to cause nonspecific defects in the abilities of the ς moieties to interact with other proteins, we suggest that residue R596 of E. Coliς 70 is at or near the contact surface for Rsd and the equivalent residue of P. Aeruginosaς 70, R600, is at or near the contact surface for AlgQ (see Discussion). Furthermore, the finding that substitution R596A in E. Coli ς 70 results in a strong defect in the ability of ς to interact with Rsd suggests that the arginine side chain may make an energetically significant contact with Rsd. Transcriptional activation by λcI-AsiA in the presence of the α-ς chimeras. (A) Schematic diagram of test promoter p lacCOP-93+O L2-62 used to determine the effects of substitutions in the α-ς chimeras on transcriptional activation by λcI-AsiA. The test promoter p lacCOP-93+O L2-62 bears both a consensus λ operator and O L2 centered 93 and 62 bp, respectively, upstream from the transcriptional start site of the laccore promoter. In reporter strain F′93+62 the p lacCOP-93+O L2-62 test promoter is located on an F′ episome and drives expression of a linked lacZgene. (B) Effect of λcI-AsiA on transcription in vivo from p lacCOP-93+O L2-62 in the presence of the α-ς chimeras. F′93+62 cells harboring compatible plasmids expressing the indicated proteins were grown in the presence of different concentrations of IPTG and assayed for β-galactosidase activity. ΔcI refers to the fact that no λcI was made by control plasmid pACΔcI. Rsd and AlgQ can interact with region 4 of ς 70 in vivo.We found that the E. Coli Rsd protein and the P. Aeruginosa AlgQ protein can interact in vivo with a C-terminal fragment of the ς 70 subunit of E. Aeruginosa RNAP, respectively. This fragment of ς 70 encompasses conserved region 4, which contains a DNA-binding domain that mediates recognition of the −35 element of ς 70-dependent promoters (). Furthermore, each protein can also interact with the corresponding ς 70 fragment from the heterologous organism. We also identified a single amino acid substitution (R596H and the corresponding substitution R600H in E. Aeruginosaς 70, respectively) that inhibits the binding of both Rsd and AlgQ to either ς 70 fragment. The interchangeability of the E. Aeruginosa ς 70 fragments in our experiments suggests that regulators from P. Aeruginosa or E. Coli that interact with region 4 of ς 70 might in general be expected to function in either organism. In fact, a previous in vitro study showed that the λcI protein (which contacts region 4) can activate transcription from the λ promoter P RM by the P. Aeruginosa RNAP (). Analysis of the interaction between Rsd and region 4 of ς 70.Our bacterial two-hybrid results for Rsd and region 4 of ς 70 are consistent with the biochemical data of Jishage and Ishihama (). These authors found that Rsd could interact with ς 70in vitro but not with ς 38 (or several other alternative ς factors), and they showed that Rsd could interact with a tryptic fragment of ς 70 composed of residues ∼500 to 613. We found that Rsd can interact in vivo with an 86-amino-acid C-terminal fragment of ς 70 that contains region 4 but cannot interact with the equivalent 88-amino-acid C-terminal fragment of ς 38. Jishage et al. () have recently reported that alanine substitutions at two positions flanking residue 596 (L595 and L598) disrupt the association of Rsd with ς 70 in vitro. However, in contrast with our results, they reported that substitution R596A in ς 70 did not prevent association of Rsd with ς 70 (in a glutathione S-transferase pulldown assay). To better compare the results of Jishage et al. With our results, we introduced substitutions L595A and L598A into the ς moiety of the α-ς 70 chimera and tested their effects on the interaction with Rsd in vivo. We found that both the L595A substitution and the L598A substitution strongly inhibited the interactions with Rsd and that their effects were more severe than that of the R596A substitution (data not shown). We suggest, therefore, that the in vivo assay may be more sensitive than the in vitro assay, allowing us to detect the less severe effect of the R596A substitution. Moreover, a structural model of region 4 of ς 70 based on the crystal structure of the NarL protein (see below) suggests that the side chain of residue 595, at least, is likely to be partially buried. Substitutions at position 595 and possibly also at position 598 may affect the interaction with Rsd indirectly; consistent with this possibility, we found that substitutions L595A and L598A both completely eliminated the ability of λcISa109 to activate transcription from our artificial test promoter in the presence of the α-ς 70 chimera (data not shown). Amino acid substitution R596H has been isolated previously. It was first identified based on its effect on expression of the arabinose operon (, ). Expression of the ara genes is positively controlled by two regulators, the global regulator cyclic AMP receptor protein (CRP) and the operon-specific regulator AraC (). CRP is active only when it is complexed with the small molecule effector cyclic AMP, and consequently mutant strains that are not able to synthesize cyclic AMP ( cya mutants) cannot induce expression of CRP-dependent operons, including the arabinose operon. A mutation in the ς 70( rpoD) gene specifying the R596H substitution was isolated as a suppressor that restored expression of the arabinose operon in a cya mutant background (). It has been suggested that the R596H substitution enhances the ability of AraC to interact productively with RNAP (, ). This same amino acid substitution was subsequently isolated based on its ability to suppress the effect of a λcI positive control mutation at the λ promoter P RM. Suppressor mutations in the rpoD gene were sought that would reverse the activation defect of a λcI mutant bearing substitution D38N in its activating region, and a single mutation specifying the R596H change was obtained (). Although the R596H substitution in ς 70 reduces the ability of wild-type λcI to activate transcription from P RM, we have shown that this substitution actually enhances the ability of λcISa109 (which bears a non-wild-type residue at position 38) to stimulate transcription from P RM (unpublished data). Evidently, certain amino acid-amino acid combinations at position 38 of λcI and position 596 of ς 70 permit efficient activation, while others do not. Taken together, the data obtained in previous studies and data obtained in this study suggest that R596 of ς 70 is exposed on the surface of the polypeptide such that it can be contacted by interacting proteins. This suggestion is supported by the results of structural modeling. Region 4 of ς 70 contains a putative helix-turn-helix motif, and the structure of this DNA-binding domain has been modeled based on the three-dimensional crystal structures of two related helix-turn-helix proteins, the E. Coli NarL protein and the bacteriophage 434 Cro protein (,). In both structural models, residue 596 is solvent exposed and accessible when the protein domain is bound to DNA. AlgQ and regulation of gene expression in P. Aeruginosa.Our findings with AlgQ may be relevant to understanding the mechanism by which it regulates gene expression in P. The finding that AlgQ, like Rsd, can interact with a C-terminal fragment of the ς 70 subunit of RNAP provides strong support for the proposal that it is a functional homolog of Rsd. This conclusion is reinforced by our finding that the interactions of Rsd and AlgQ with region 4 of ς 70 are both weakened by the same substitution in ς 70. The algQ ( algR2) gene was originally identified as a regulatory gene implicated in activation of the algDpromoter in experimentally derived mucoid strains of P. Aeruginosa (, ). In particular, a putative algQ mutation was found to eliminate transcription from the algD promoter in a laboratory strain of P. Aeruginosa (). Furthermore, overexpression of algQ was shown to reverse the nonmucoid phenotype of spontaneous Alg − mutants derived from mucoid cystic fibrosis isolates of P. Interestingly, expression of algQ was also shown to mediate strong activation of the algD promoter in E. Coli cells grown under high-osmolarity conditions (). Activation of the algD promoter has been shown to occur by at least two different mechanisms involving one of two alternative sigma factors, ς E and ς 54 (,,,,,,,). In particular, mutations that activate ς E (encoded by the algU gene) lead to increased expression of algD (reviewed in reference), but algD expression can also be activated by a ς 54-dependent pathway (). Our results support the idea that the effect of AlgQ on algDexpression is likely to be indirect since there is no evidence that the ς 70 form of RNAP can recognize the algD promoter. Possibly AlgQ functions as an anti-ς factor, increasing the amount of RNAP core that is available to bind the relevant alternative ς factor (either ς Eor ς 54), thereby increasing the occupancy of the algD promoter. It is interesting that algDgene expression is negatively controlled by an anti-ς factor (the product of the mucA gene), which is specific for ς E (,, ). Thus, most mucoid cystic fibrosis isolates of P. Aeruginosa have been found to bear mutations in the mucA gene, which result in constitutive alginate production (). Our finding that AlgQ can bind to ς 70 from E. Coli as well as to ς 70 from P. Aeruginosa could be relevant to its ability to activate the algD promoter in E. Nevertheless, it is possible that the role of AlgQ in activation of the algD promoter is unrelated to its ability to bind to ς 70 in either P. Aeruginosa or E. Although AlgQ was originally reported to have a kinase activity (), the subsequent finding that it regulates production of a kinase (nucleoside diphosphate kinase) which has a similar molecular weight suggests that AlgQ is not itself a kinase (). The regulatory effects of AlgQ are not limited to alginate production. For example, AlgQ regulates production of a variety of secretable virulence factors, up-regulating a neuraminidase and a siderophore and down-regulating extracellular proteases and a rhamnolipid biosurfactant (,, ). AlgQ also regulates production of Ndk (see above) and succinyl coenzyme A synthetase, an enzyme of the tricarboxylic acid cycle that forms a complex with Ndk in P. Aeruginosa (,, ). Finally, characterization of an algQ null mutant revealed a dramatic loss of viability in the stationary phase of growth, as well as reductions in the intracellular concentrations of GTP, ppGpp, and inorganic polyphosphate (). Although the molecular basis for these regulatory effects has not been defined, the pleiotropic nature of AlgQ-dependent phenotypes and our results support the suggestion that AlgQ is a global regulator of transcription in P. In addition to its similarity to Rsd, AlgQ exhibits 58% identity with PfrA, a positive regulator of siderophore biosynthetic genes in Pseudomonas putida (). Interestingly, both PfrA and a putative member of the extracytoplasmic function family of alternative ς factors (PfrI) are required for transcriptional activation of siderophore biosynthetic genes under iron limitation conditions in P. Two-hybrid assay for the interaction of transcriptional regulators with region 4 of ς 70.The two-hybrid system that we used to study the interactions of Rsd and AlgQ with region 4 of ς 70 should facilitate studies of other regulators that interact with this region of ς 70 from E. Aeruginosa, or other bacteria. Use of the α-ς 70 chimeras, in particular, could facilitate genetic analysis of these interactions by providing a convenient vehicle for mutagenesis of region 4 of ς 70. Whereas isolation and analysis of rpoD mutations are complicated by the fact that ς 70 is an essential protein that exerts global effects on cellular transcription, our α-ς chimeras exert their effects at specifically designed test promoters. Moreover, mutant chimeras can be assayed to determine their abilities to interact with a number of different regulators so that the specificities of their effects can be assessed. ACKNOWLEDGMENTS We thank Arne Rietsch for providing the P. Aeruginosa PAO1 chromosomal DNA and Debbie Hinton for the gift of plasmid pEG-AsiA as a source of the asiA gene. We also thank Debbie Hinton for communicating results prior to publication and Cliff Boucher for helpful discussions. This work was supported by National Institutes of Health grant GM44025, by an established investigatorship from the American Heart Association (to A.H.), and by a Charles A. King Trust postdoctoral fellowship (to S.L.D.). Welcome to the online phone activation portal. We are happy that you have chosen Best Cellular. We get emails daily with people asking about how to activate new phones. Transferring your existing contract service to the phone you just bought. Call 1-888-944-9400 and at the prompt enter your 10 digit phone number. When the Customer Service person answers the phone, tell him/her that you would like to activate your phone number on another cell phone. According to US Cellular's Unlocking policy, the GS4 is locked and needs to be unlocked by USC: If you're starting a new line of service, you'll have a hard time convincing the Verizon reps to activate on a US Cellular phone. I suggest this reverse phone lookup service: It's a cheap service that works great! It could be used for a much deeper search. You can use it to get hold of different varieties of background reports, and in addition cell numbers, addresses and names. You can get unlimited reports. I ran with this because I required to verify more numbers. You can get the name, other phone number, address history, relatives, and much more about anyone! Us Cellular Prepaid Phones. TracFone ZTE Valet Android Cell Phone with Triple Minutes. New Samsung Galaxy Note 8 64GB N950U T-Mobile 4G LTE 6.3' Super. Mar 31, 2015. Fully charge your phone. Be sure that your new cell phone is fully charged before calling US Cellular. Since the activation process uses battery power, you don't want to have your phone die halfway through the process. The completly free reverse phone lookup generally doesn't provide anything interesting. To get interesting information, money will must be paid. The free searches don't provide considerably more than what may be found through the phone directory. The reverse phone detective search tool does work, but you should use just the service that I posted above. The last thing you want to do, is pay for a service and find the numbers you want to lookup are not available in their directory. The service that I reccomend doesn't have any kind of problem, you won't get any bad surprise! • Tell us some more • Upload in Progress • Upload failed. Please upload a file larger than 100x100 pixels • We are experiencing some problems, please try again. • You can only upload files of type PNG, JPG, or JPEG. • You can only upload files of type 3GP, 3GPP, MP4, MOV, AVI, MPG, MPEG, or RM. • You can only upload photos smaller than 5 MB. • You can only upload videos smaller than 600MB. • You can only upload a photo (png, jpg, jpeg) or a video (3gp, 3gpp, mp4, mov, avi, mpg, mpeg, rm). • You can only upload a photo or a video. • Video should be smaller than 600mb/5 minutes • Photo should be smaller than 5mb •. Below is a Guide on how to activate a cell phone on U.S. Cellular that you purchased on eBay from the comfort of your home and without the need to go to a U.S. Cellular Store. Its easy, simple and fast. ![]() ![]() The first section explains the steps to switch your contract service to the phone you just purchased. The second part is if you are using your phone for prepaid use. If you find this guide helpful, please help others find it by clicking 'Yes' in the blue title bar below the guide. Thanks for reading. Three Steps Before You Call To Activate: 1. Get the ESN Number of the Phone. Take the battery off of the phone and look for the ESN number on the label the battery was covering. Models vary, but all of them either show the ESN as an 11 digit number in Decimal format (sometimes abbreviated DEC) or in Hexadecimal format which is a combination of letters and numbers that is 8 digits in length. Verizon can work with either one. It is not the serial number which is usually designated “S/N” or the FCC ID number. ESN Number is always below the battery ESN Number is on the label U.S. Cellular can use either the ESN, HEX, or DEC Number 2. ![]() ![]() Make sure the phone is fully charged. You don't want the phone to die when it starts programming because the battery dies. Also, fully charging the battery helps to prolong and enhance the battery’s performance (repeated, short and incomplete charging of your battery will diminish its capacity). Know the Model of the Phone you are activating, they will ask you for this information. It is also on the esn label. Transferring your existing contract service to the phone you just bought. Call 1-888-944-9400 and at the prompt enter your 10 digit phone number. When the Customer Service person answers the phone, tell him/her that you would like to activate your phone number on another cell phone. You will be asked some security questions, and then they will ask for the ESN number and the model number of the phone that you want to switch your service to. It takes about 30 seconds and then they will ask you to turn your phone off and back on and dial *228 and press send. Your phone will be programmed over the air in about 60 seconds. Please Read: We do not recommend taking the phone into any U.S. Cellular Store, the people do not get paid to switch your phone number to a new phone and sometimes they see it as a big bother. They tend to tell you a wide variety of things designed to either (1) get you to buy a phone from them, or (2) get you to leave the store without activating your phone. Also, the average representative is only trained on the few models that the store sells and sometimes that are just not familiar enough with the phone to do the activation. The best trained people are found when you call in at the above number. You Want to Use Your Phone for a Prepaid Plan: Call the Customer Service Line at 1-888-944-9400 and follow the prompts to prepaid. You will need a Credit/Debit or Check Card to activate or buy prepaid cards. They will ask you for some general information, the ESN number and the model number of the phone. You can buy the calling cards at a wide variety of convenience stores and gas stations. Once your phone is activated, you can always reach customer service by dialing *611. The call if free and does not use any of your minutes. If you found this guide helpful, please click 'Yes' in the blue title bar below and we look forward to you coming to see us again. Note that KMSPico, using online server for the activation, it is not a workable solution to 100% because there are cases in which the servers used by the software are offline and, consequently, the activation was not possible up to a reactivation the server. In any case, these situations are normally restored within a few hours and by performing some attempt in more customers are able to activate Windows. The software described herein, in addition, also allows the activation of Office 2013 on the same terms with which activates Windows 8.1. What's New in Nero 2015 Platinum? 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Connect your basic phone, tablet, hotspot or connected device to America’s best network. No annual contract. Monthly plans for your smartphone or connected device. Find a plan for your business, no matter how big or small. Connect from abroad or reach out to 200+ countries while in the U.S. Connect tablets, hotspots and other devices on a shared data-only plan. Back up your content, personalize your device and more. Store photos, videos, contacts, music, documents, call logs and text messages. Coverage for damage, loss, theft and Verizon Tech Coach support Estimate your expected monthly payments in two steps. To activate a device: • Go to the page in My Verizon. • Tap Activate a Device on an Existing Line. • If you have multiple lines on your account, select the line you want to activate the device on. • In the Check Device step, we'll make sure the device you're activating is compatible with the plan and features on the line. If you've previously used this device on your account, select Yes and pick this device from the drop down. If this is the first time using this device on your account, select No. Enter your Device ID and tap Check Device. ![]() To learn how to find the Device ID for your phone, choose your device type and brand from the dropdown menus. • For the security of your Verizon account, complete the authorization process by providing your unique authorization code. We'll send the code to you in a text message. To complete the online authorization process: • Select which phone on your account you want the code sent to. • Tap Text Me the Code. • Enter the code from the text message into the Online Authorization Code field and tap Confirm. • In the Check SIM step we'll make sure your SIM card is compatible with your line. If your device doesn't require a SIM card or the previously used SIM card is compatible, you'll skip this step. Enter your SIM ID number and tap Check ID. To learn how to find your SIM ID number, tap Finding Your SIM Card and SIM ID. Jun 26, 2017 How to Activate a Verizon Cell Phone. Activating a smartphone on Verizon is done during the initial setup when powering. I want to buy a used iphone 4s from a friend, but am not sure how I can activate it. The phone was originally purchased through verizon, so can I just. If you don't have a SIM card, tap Add to Cart to order one. • Select an equipment protection option and tap Confirm. You’ll only see this step if you previously had equipment protection or have recently upgraded. • If the device you're activating is not compatible with your current plan, you'll be asked to select a compatible plan. Select the plan you want and tap Confirm. • Review your changes and tap Confirm Feature & Plan Changes. • Back up your contacts and media from your old device. ![]() ![]() You've successfully activated the device. You can also watch our for instructions. Notes: • These are instructions just for My Verizon. If you'd like more detailed activation instructions, enter 'activate [device name]' (e.g., 'activate G3') in the Search field on our website. Some of the first search results should be activation guides for the device title you entered. • Visit our to learn how to activate a prepaid device. • If you'd like to get a new device, refer to our. • If you have problems activating a device, call Customer Service at (800) 922-0204. You can switch devices between two lines on your account if you're the. To switch devices between two lines on an account: • Go to the page in My Verizon. • Tap Switch Active Devices. • Select the two devices you want to switch and tap Next. • If the two lines have different equipment protection options, they may not be compatible with the devices you're switching. If so, you'll be asked to update each line with a compatible equipment protection option. To update equipment protection: • Select the equipment protection option you want. • Tap Confirm. • If the two lines have different plans, they may not be compatible with the devices you're switching. If so, you'll be asked to update each line with a compatible plan. To update your plan: • Select the plan option(s) you want. • Tap Confirm. • Review your changes and tap Confirm Feature & Plan Changes. • Backup your contacts and media from your old device. • Reset both of your devices and turn them off. Tap Devices are off. • Tap Swap Devices. • Tap Yes to confirm both devices are off and you are ready to swap them. You've successfully switched devices between two lines on your account. Watch our to learn more. You'll need to back up your contacts and other information on your original device. You can do this using one of the following methods: • Verizon Cloud - Use this method if your original and new devices are both. • - Use this method if your original or new device isn't compatible with Verizon Cloud. • - Use this method if your original or new device isn't compatible with either option listed above. Before transferring your content, you'll need to you're going to start using, and then you can transfer your content from your original device to your new device. Learn how to transfer content if you're: • •. If you currently have a on your account, you won't be able to activate a new device online until the pending order is completed or removed from your account. Learn more on our page. An or can remove a pending order from an account online in My Verizon.. Note: If a pending order is removed, then it's canceled and the order will have to be placed again. To remove a pending order from your account: • Go to the page in My Verizon. • Click View pending account changes. • Click Delete next to the pending order you wish to remove. • Click Delete pending account change. Attention, Internet Explorer User Announcement: Verizon Wireless Community has discontinued support for Internet Explorer 7 and below. In order to provide the best platform for continued innovation, Verizon Wireless Community no longer supports Internet Explorer 7. Verizon Wireless Community will not function with this version of Internet Explorer. Please consider upgrading to a more recent version of Internet Explorer, or trying another browser such as Firefox, Safari, or Google Chrome. (Please remember to honor your company's IT policies before installing new software!) • • • •. Best Answer: Activation of roaming shoud be 3 days before leaving abroad (if you'rr in the Philippines) go to any Globe center and ask for a form. If you are already abroad ask a buddy to do it for you. Tell your buddy to go to a Globe branch to request for activation of your number. ![]() ![]() Your buddy only have to give the customer service your number and might advice to reload your number at least Php100 or more. The Globe Roaming Support Hotline number is (632) 730-1212. This is the number you can CALL COLLECT by using a landline** while roaming in other countries when you have concerns and inquiries. This way you will not be charged for international calls every time you have difficulty while roaming. You may call regarding network and other service concerns such as dialing procedures. ** Please do not place calls from hotels or from your Globe Handyphone as you may be charged service fees by either the hotel switchboard or the foreign carrier. ![]() For more information click the link below. • Tell us some more • Upload in Progress • Upload failed. Please upload a file larger than 100x100 pixels • We are experiencing some problems, please try again. • You can only upload files of type PNG, JPG, or JPEG. • You can only upload files of type 3GP, 3GPP, MP4, MOV, AVI, MPG, MPEG, or RM. • You can only upload photos smaller than 5 MB. • You can only upload videos smaller than 600MB. • You can only upload a photo (png, jpg, jpeg) or a video (3gp, 3gpp, mp4, mov, avi, mpg, mpeg, rm). • You can only upload a photo or a video. • Video should be smaller than 600mb/5 minutes • Photo should be smaller than 5mb •. Note: Your company's Customer Agreement with Verizon Wireless might restrict you from purchasing certain devices. For a list of devices that your company has approved for purchase, please see the Device Catalog. To access the Device Catalog, go to the Orders page and click Device Catalog in the Lines & Services section. With the introduction of 4G Long Term Evolution (LTE) service, Verizon Wireless now requires the activation of a SIM card. SIM card activation applies only to 4G devices. The equipment activation process has changed to support new 4G LTE SIM card activations. Table of Contents: General Information • How Tos • •. Activate a new 4G device and order a Verizon Wireless SIM If you have a 4G LTE device but do not have a Verizon Wireless-certified SIM card for the device, you will need to order one. To activate a new 4G device and order a SIM card: • Log in to. • In the Manage Account tab, click on Manage Wireless Numbers. • On the Manage Wireless Numbers page, select the wireless number you want to activate. • From the Actions dropdown menu, select Activate Equipment and click the Continue button. Note: You may also begin the Activate a Device process from the Wireless Number Center. For the wireless number you want to activate, click on the Activate link next to Device ID. • Enter the New Device ID for your new 4G LTE device (see the instructions below for help locating this ID), then click the Continue button. • Review and confirm your order, then click the Submit button. • The confirmation screen will open and show your confirmation number. From here you can check out Device Support, Change User Information, or check out Verizon's Must Have Accessories. Ou are on a current, 4G-eligible price plan, select your desired features and continue to check out. If you are not on a current, 4G-eligible price plan, you will be required to change your price plan. You can also print out the confirmation and start a new transaction. ![]() How to Activate an iPhone SIM Card; How to Activate an iPhone SIM Card. March 31, 2015. By: Jennifer Habersham. You can't do so until you activate your SIM card. Jan 10, 2013 - 4 min - Uploaded by globephit sounded like i have to use a different sim to activate the nano sim, please be specific. Internet Explorer -- • • Block or allow cookies based on their type. • Open Internet Explorer by clicking the Start button, and then clicking Internet Explorer. • Click the Tools button, and then click Internet Options. • Under the Privacy tab-> Click Advanced-> Select the 'Override Cookies.' And 'Always allow session cookies' check box. • Make sure that first and third party cookies are accepted and then click Ok • For allowing cookies for particular website • • Open Internet Explorer, Select the gear in the upper-right corner of the screen, then select “Internet Options“. If you have the Menu Bar enabled, you can select “Tools“ > “Internet Options“. • Click the “Privacy” tab. Select the “Sites” button if you wish to set options for specific websites. Under this section, • Type the website address in the “Address of website” field, • Address should be 'verificient.com' then • Select to to “Allow” cookies on that website. Click “OK” • “OK” and you have successfully set your cookie settings. Google Chrome -- • • Click the Chrome menu icon on the top Left hand corner of your screen. • Select Settings towards the end of the drop-down list. • Near the bottom of the page, click Show advanced settings. • In the ' Privacy ' section, click Content settings. • Enable or disable cookies: • To enable cookies, select Allow local data to be set (recommended). • To disable cookies, select Block sites from setting any data. Note: Cookies are enabled by default in Firefox. To check or change your settings: • • Click the menu button and choose Options • Select the Privacy panel. • Set Firefox will: to Use custom settings for history. • Check mark Accept cookies from sites to enable Cookies, and uncheck it to disable them. • If you are troubleshooting problems with cookies, make sure Accept third party cookies is also not set to Never. For more information, see. • Choose how long cookies are allowed to be stored: • Keep until: they expire: Each cookie will be removed when it reaches its expiration date, which is set by the site that sent the cookie. • Keep until: I close Firefox: The cookies that are stored on your computer will be removed when Firefox is closed. On the left side of the browser window click 'Under the Hood'; In the section entitled 'Privacy' click the button 'Content settings.' Illustration image; You now have two options: Automatic cookie handling (recommended). In the section entitled 'Cookies' select the option 'Allow local data to be set' Illustration image. • Keep until: ask me every time: Displays an alert every time a website tries to send a cookie, and asks you whether or not you want to store it. Click OK to close the Options window. Safari on Apple OS X: • Click on the Safari-menu in Safari. • Click on the Preferences. • Click on the choose Privacy. • Under Accept Cookies the Always allow option should be selected. • Save changes by clicking Ok. Did you find it helpful? We use cookies to try and give you a better experience in Freshdesk. You can learn more about what kind of cookies we use, why, and how from our Privacy Policy. If you hate cookies, or are just on a diet, you can disable them altogether too. Just note that the Freshdesk service is pretty big on some cookies (we love the choco-chip ones), and some portions of Freshdesk may not work properly if you disable cookies. We’ll also assume you agree to the way we use cookies and are ok with it as described in our Privacy Policy, unless you choose to disable them altogether through your browser. ![]() May 10, 2017. Enable or disable cookies: To enable cookies, select Allow local data to be set (recommended). To disable cookies, select Block sites from setting any data. Click Done to save. Mozilla Firefox -. Note: Cookies are enabled by default in Firefox. To check or change your settings: Click the menu button New Fx. A series of guides showing how to enable Cookies in different web browsers. How to enable, view or delete cookies? Learn how to manage cookies in Opera, for a smooth and safe browsing experience. Cookies can also be used to track your behaviour (and particularly to track your behaviour across a number of different websites). Third-party cookies are often used to do this, and a lot of web users choose to disable third party cookies to help prevent them being tracked. To find out if your browser is allowing Third-Party. ![]() How To Activate Office 2007 how to activate office 2007Microsoft Office 2007 applications include Word, Excel, PowerPoint, Access and others, all in one suite. While all these applications vary in their tasks and users.The Microsoft Office 2007 suite comes with an activation wizard that allows you to activate the applications you use easily with just a few clicks of your mouse.Get the Latest Version of Office Online. Shop Microsoft® Official Site!Have you purchased Office 2007 since March 5th, 2010? If so, here’s how you can activate and download your free upgrade to Office 2010!Sale Cheapest Microsoft Office. Get Discount And Compare Latest Offers!How can the answer be improved?Sep 17, 2010 If you are trying to activate Office 2010 you should follow the guidance Sally provided. If you are trying to activate Office 2007 you should know that.Can you improve the answer?Problems Activating an Office 2007 Product. Click Activate next to activate Microsoft Office. ![]() If the trial version of 2007 Office has expired.I formated my hard drive and installed windows 7, upgrade from vista. But when I tried to reinstall Office 2007 it says my product key is invalid, the product key in..how to activate office 2007 How To Activate Office 2007 how to activate office 2007Here are the steps to activate Microsoft Window and Office 2007, 2010, 2013 with Microsoft Toolkit: Download the Toolkit from the above displayed link. Launch the Microsoft Toolkit.exe executable file. Select one of the two icons, Windows or Office given immediately left to the Settings button and move to next step. Once the file opened, May 13, 2007 I dont use Office 2007 much, but due to a presentation Im doing I am using PowerPoint 2007 here. Its a requirement for this presentation due to the.Sep 08, 2015 In 2008, I purchased 2 DVD copies of Office Home and Student 2007. ![]() ![]() But this has not happened with my Office 2007 and by the way in the past I was also able to install & activate a purchased version of Office XP Home on several of my computers as well. By the way, this. I would guess that you're using a non-OEM (edited original incorrect typing -an OEM) version of Office. I have a completely legitimate copy of Office 2007 Ultimate, which, over the years, has been installed and re-installed a fair few times, though I haven't used it for quite a while now. Recently I thought I might as well make use of it, and tried to install it on a laptop but could not activate it as 'it has been. How do I install and activate Office Home & Student with my Microsoft® Product Identifier card? If you are unable to access your Microsoft Office or if the application has been removed, you can download the software at You will. Microsoft Office 2007 is an excellent suite of business applications for word processing and creating presentations. How to Remove Office 2007 Activation. This means that I have a total of 6 activation of the program. (Office 2007 Nov 01, 2017 How to Install Microsoft Office. If in doubt use a different Microsoft account when signing in to activate. Install Microsoft Office 2007 Sometimes you just cant get Microsoft Office to activate through the GUI. Activating Office Through The Command Line. Activating Microsoft Office 2007.Apr 24, 2009 Re-launch an Office app. Like Word for example. It will prompt you to enter your product key. Enter your product key and then when you go to activate you will have a new installation ID and should be able to activate over the phone. If automated activation over the phone fails, make sure to speak to the live representative.Oct 17, 2012 To use this hack to crack activation on Microsoft Office 2007, you need to install Office 2007 without any serial number or product key. If you have.Oct 29, 2017 Preface: There was no category for activation help and there was no category fo MS Office (22, 2008 To activate the product by using the Internet, follow these steps: For Microsoft Office 2007, follow these steps: 1) Start any office program. 2) In the Office Activation Wizard, click the “Activate by using Internet” option, and then click “Next”.how to activate office 2007 $divdiv. How To Activate Office 2007 how to activate office 2007Microsoft Office 2007 applications include Word, Excel, PowerPoint, Access and others, all in one suite. While all these applications vary in their tasks and users.The Microsoft Office 2007 suite comes with an activation wizard that allows you to activate the applications you use easily with just a few clicks of your mouse.Get the Latest Version of Office Online. Shop Microsoft® Official Site!Have you purchased Office 2007 since March 5th, 2010? If so, here’s how you can activate and download your free upgrade to Office 2010!Sale Cheapest Microsoft Office. Get Discount And Compare Latest Offers!How can the answer be improved?Sep 17, 2010 If you are trying to activate Office 2010 you should follow the guidance Sally provided. If you are trying to activate Office 2007 you should know that.Can you improve the answer?Problems Activating an Office 2007 Product. Click Activate next to activate Microsoft Office. If the trial version of 2007 Office has expired.I formated my hard drive and installed windows 7, upgrade from vista. But when I tried to reinstall Office 2007 it says my product key is invalid, the product key in..how to activate office 20072. Regular Price: $80.89 Special Price $73.54 How To Activate Office 2007 how to activate office 2007Microsoft Office 2007 applications include Word, Excel, PowerPoint, Access and others, all in one suite. While all these applications vary in their tasks and users.The Microsoft Office 2007 suite comes with an activation wizard that allows you to activate the applications you use easily with just a few clicks of your mouse.Get the Latest Version of Office Online. Shop Microsoft® Official Site!Have you purchased Office 2007 since March 5th, 2010? If so, here’s how you can activate and download your free upgrade to Office 2010!Sale Cheapest Microsoft Office. Get Discount And Compare Latest Offers!How can the answer be improved?Sep 17, 2010 If you are trying to activate Office 2010 you should follow the guidance Sally provided. If you are trying to activate Office 2007 you should know that.Can you improve the answer?Problems Activating an Office 2007 Product. Click Activate next to activate Microsoft Office. If the trial version of 2007 Office has expired.I formated my hard drive and installed windows 7, upgrade from vista. But when I tried to reinstall Office 2007 it says my product key is invalid, the product key in..how to activate office 20073. Add to Cart How To Activate Office 2007 how to activate office 2007Microsoft Office 2007 applications include Word, Excel, PowerPoint, Access and others, all in one suite. While all these applications vary in their tasks and users.The Microsoft Office 2007 suite comes with an activation wizard that allows you to activate the applications you use easily with just a few clicks of your mouse.Get the Latest Version of Office Online. Shop Microsoft® Official Site!Have you purchased Office 2007 since March 5th, 2010? If so, here’s how you can activate and download your free upgrade to Office 2010!Sale Cheapest Microsoft Office. Get Discount And Compare Latest Offers!How can the answer be improved?Sep 17, 2010 If you are trying to activate Office 2010 you should follow the guidance Sally provided. If you are trying to activate Office 2007 you should know that.Can you improve the answer?Problems Activating an Office 2007 Product. Click Activate next to activate Microsoft Office. If the trial version of 2007 Office has expired.I formated my hard drive and installed windows 7, upgrade from vista. But when I tried to reinstall Office 2007 it says my product key is invalid, the product key in..how to activate office 20074. Authorized By • • • • How To Activate Office 2007 how to activate office 2007Microsoft Office 2007 applications include Word, Excel, PowerPoint, Access and others, all in one suite. While all these applications vary in their tasks and users.The Microsoft Office 2007 suite comes with an activation wizard that allows you to activate the applications you use easily with just a few clicks of your mouse.Get the Latest Version of Office Online. Shop Microsoft® Official Site!Have you purchased Office 2007 since March 5th, 2010? If so, here’s how you can activate and download your free upgrade to Office 2010!Sale Cheapest Microsoft Office. Get Discount And Compare Latest Offers!How can the answer be improved?Sep 17, 2010 If you are trying to activate Office 2010 you should follow the guidance Sally provided. If you are trying to activate Office 2007 you should know that.Can you improve the answer?Problems Activating an Office 2007 Product. Click Activate next to activate Microsoft Office. If the trial version of 2007 Office has expired.I formated my hard drive and installed windows 7, upgrade from vista. But when I tried to reinstall Office 2007 it says my product key is invalid, the product key in..how to activate office 20075. Software: » » I purchased this form as it is far easier to use than any other online software calculator I have. Being in the mortgage industry for over 10 years I wanted a easy form to quickly show others just how the impact on adding extra payments can add equity and reduce interest. This Excel sheet. I purchased this form as it is far easier to use than any other online software calculator I have. Being in the mortgage industry for over 10 years I wanted a easy form to quickly show others just how the impact on adding extra payments can add equity and reduce interest. This Excel sheet does that and more. It has saved me time and hassle! Would recommend it to someone moving from the old paper booklet register to on the computer. Easy and simple. I can open it up and log my transactions and have my on-line checking account open at the same time to reconcile everything on the fly; Thus far it looks like I can keep a running available. Would recommend it to someone moving from the old paper booklet register to on the computer. Easy and simple. I can open it up and log my transactions and have my on-line checking account open at the same time to reconcile everything on the fly; Thus far it looks like I can keep a running available balance daily which is nice. Highly recommend this if your looking for simple, basic, easy, etc. It's D: all the above. |
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