Since its development in 1999, plasmid-based reverse genetics has been effectively applied to numerous aspects of influenza studies which include revolutionizing the production of seasonal and pandemic influenza vaccine seed strains. We recommend downloading the newest version of Flash here, but we support all versions 10 and above. Older browsers that do not support HTML5 and the H.264 video codec will still use a Flash-based video player. This volume is a compilation of sixteen chapters that detail reverse genetics protocols. (gross), © 2020 Springer Nature Switzerland AG. price for Bulgaria Virology 198:415–426, Palese P, Zheng H, Engelhardt OG, Pleschka S, Garcia-Sastre A (1996) Negative-strand RNA viruses: genetic engineering and applications. We use cookies to enhance your experience on our website. Avian Dis 49:207–213, © Springer Science+Business Media New York 2014. This volume is a compilation of sixteen chapters that detail reverse genetics protocols. We may use this info to send you notifications about your account, your institutional access, and/or other related products. Springer is part of, Please be advised Covid-19 shipping restrictions apply. Not logged in Your access has now expired. Virol J 4:102, Song MS, Baek YH, Pascua PN, Kwon HI, Park SJ, Kim EH, Lim GJ, Choi YK (2013) Establishment of Vero cell RNA polymerase I-driven reverse genetics for Influenza A virus and its application for pandemic (H1N1) 2009 influenza virus vaccine production. Reverse genetics is the creation of a virus from a full-length cDNA copy of the viral genome, referred to as an “infectious clone,” and is one of the most powerful genetic tools in modern virology. Please review prior to ordering, Includes cutting-edge methods and protocols, Provides step-by-step detail essential for reproducible results, Contains key notes and implementation advice from the experts, ebooks can be used on all reading devices, Institutional customers should get in touch with their account manager, Usually ready to be dispatched within 3 to 5 business days, if in stock, The final prices may differ from the prices shown due to specifics of VAT rules. A subscription to JoVE is required to view this content.You will only be able to see the first 20 seconds. CYBER DEAL: 50% off all Springer eBooks | Get this offer! Reverse genetics is the creation of a virus from a full-length cDNA copy of the viral genome, referred to as an “infectious clone,” and is one of the most powerful genetic tools in modern virology. Simplified Reverse Genetics Method to Recover Recombinant Rotaviruses Expressing Reporter Proteins, Unable to load video. A JoVE representative will be in touch with you shortly. Learn more. Food Animal Health Research Program, Ohio Agricultural Research and Development Center, https://doi.org/10.1007/978-1-4939-0758-8_4. Proc Natl Acad Sci U S A 96:9345–9350, Fodor E, Devenish L, Engelhardt OG, Palese P, Brownlee GG, Garcia-Sastre A (1999) Rescue of influenza A virus from recombinant DNA. If that doesn't help, please let us know. J Virol Methods 193:603–610, Massin P, Rodrigues P, Marasescu M, van der Werf S, Naffakh N (2005) Cloning of the chicken RNA polymerase I promoter and use for reverse genetics of influenza A viruses in avian cells. Influenza Other Respir Viruses 7:249–256, Boyer J-C, Haenni A-L (1994) Infectious transcripts and cDNA clones of RNA viruses. Although the reverse genetics system has been utilized in many viruses, the system using a helper virus was developed for rotavirus in 2006. Please check your Internet connection and reload this page. (Ed.). Cold Spring Harbor Laboratory Press, Plainview, NY, Hoffmann E, Krauss S, Perez D, Webby R, Webster RG (2002) Eight-plasmid system for rapid generation of influenza virus vaccines. If you want more info regarding data storage, please contact gdpr@jove.com. By continuing to use our website or clicking “Continue”, you are agreeing to accept our cookies. Part of Springer Nature. J Virol 73:9679–9682, Neumann G, Kawaoka Y (2002) Synthesis of influenza virus: new impetus from an old enzyme, RNA polymerase I. Reverse genetics systems have truly transformed our ability to manipulate and study negative-strand RNA … Arch Virol 158:2049–2058, Mostafa A, Kanrai P, Ziebuhr J, Pleschka S (2013) Improved dual promotor-driven reverse genetics system for influenza viruses. Reverse Genetics of RNA Viruses: Methods and Protocols guides readers through comprehensive protocols on RNA viruses, that were the most challenging to obtain and/or that were developed most recently. If the problem continues, please, An unexpected error occurred. 50.62.23.88. Coupled with in vitro mutagenesis, reverse genetics can be applied widely to accelerate progress in understanding the influenza virus life cycle, the generation of customized vaccine seed strains, development of live-attenuated vaccines, and the use of influenza virus as vaccine and gene delivery vectors. J Virol 79:13811–13816, Wang Z, Duke GM (2007) Cloning of the canine RNA polymerase I promoter and establishment of reverse genetics for influenza A and B in MDCK cells. Reverse genetics is the process of generating an RNA virus from a cDNA copy. Copyright © 2020 MyJoVE Corporation. JavaScript is currently disabled, this site works much better if you ...you'll find more products in the shopping cart. Although continual improvement in reverse genetics system is being made in different laboratories for the efficient rescue of the influenza virus, the basic concept of synthesizing viral RNA using RNA polymerase I remains the same. Reverse genetics is a method in molecular genetics that is used to help understand the function(s) of a gene by analysing the phenotypic effects caused by genetically engineering specific nucleic acid sequences within the gene. Not affiliated Springer Nature is developing a new tool to find and evaluate Protocols. Perez, Daniel R. © 2020 Springer Nature Switzerland AG. Over 10 million scientific documents at your fingertips. Generation of recombinant rotaviruses from plasmid DNA provides an essential tool for the study of rotavirus replication and pathogenesis, and the development of rotavirus expression vectors and vaccines. Vaccine 22:3175–3181, Lee CW, Lee YJ, Senne DA, Suarez DL (2006) Pathogenic potential of North American H7N2 avian influenza virus: a mutagenesis study using reverse genetics. Virology 353:388–395, Lee CW, Senne DA, Suarez DL (2006) Production of reference antisera against 15 hemagglutinin subtypes of influenza virus by DNA vaccination of chickens. If you would like to continue using JoVE, please let your librarian know as they consider the most appropriate subscription options for your institution’s academic community. To learn more about our GDPR policies click here. If the problem continues, please. … ... PCR -based protocol , infectious -subgenomic-amplicons method and systems 22 using a … The authors would like to thank Dr. Gerd Hobom (Institut fur Mikro- und Molekularbiologie, Giessen, Germany) for kindly providing pHH21 vector and Dr. Yoshihiro Kawaoka (University of Wisconsin) for providing plasmids for A/WSN/33 (H1N1) reverse genetics. enable JavaScript in your browser. Vet Microbiol 135:239–247, Lee CW, Senne DA, Suarez DL (2004) Generation of reassortant influenza vaccines by reverse genetics that allows utilization of a DIVA (Differentiating Infected from Vaccinated Animals) strategy for the control of avian influenza.

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