{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,27]],"date-time":"2026-08-27T00:39:32Z","timestamp":1787791172767,"version":"build-2784847793"},"reference-count":64,"publisher":"Public Library of Science (PLoS)","issue":"1","license":[{"start":{"date-parts":[[2015,1,23]],"date-time":"2015-01-23T00:00:00Z","timestamp":1421971200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"DOI":"10.1371\/journal.pone.0116917","type":"journal-article","created":{"date-parts":[[2015,1,23]],"date-time":"2015-01-23T20:10:16Z","timestamp":1422043816000},"page":"e0116917","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":26,"title":["Efficient Generation of Recombinant Influenza A Viruses Employing a New Approach to Overcome the Genetic Instability of HA Segments"],"prefix":"10.1371","volume":"10","author":[{"given":"Ahmed","family":"Mostafa","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pumaree","family":"Kanrai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Henning","family":"Petersen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sherif","family":"Ibrahim","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Silke","family":"Rautenschlein","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephan","family":"Pleschka","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2015,1,23]]},"reference":[{"issue":"1","key":"ref1","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1099\/0022-1317-72-1-103","article-title":"Antigenic characterization of the nucleoprotein and matrix protein of influenza C virus with monoclonal antibodies","volume":"72","author":"K Sugawara","year":"1991","journal-title":"J Gen Virol"},{"key":"ref2","doi-asserted-by":"crossref","first-page":"1861","DOI":"10.1098\/rstb.2001.0999","article-title":"The evolution of human influenza viruses","volume":"356","author":"AJ Hay","year":"2001","journal-title":"Philos Trans R Soc Lond B Biol Sci"},{"key":"ref3","doi-asserted-by":"crossref","first-page":"4269","DOI":"10.1073\/pnas.1116200109","article-title":"A distinct lineage of influenza A virus from bats","volume":"109","author":"SX Tong","year":"2012","journal-title":"Proceedings of the National Academy of Sciences of the United States of America"},{"key":"ref4","doi-asserted-by":"crossref","first-page":"e1003657","DOI":"10.1371\/journal.ppat.1003657","article-title":"New world bats harbor diverse influenza A viruses","volume":"9","author":"S Tong","year":"2013","journal-title":"PLoS Pathog"},{"key":"ref5","doi-asserted-by":"crossref","first-page":"2814","DOI":"10.1128\/JVI.79.5.2814-2822.2005","article-title":"Characterization of a novel influenza A virus hemagglutinin subtype (H16) obtained from black-headed gulls","volume":"79","author":"RA Fouchier","year":"2005","journal-title":"J Virol"},{"key":"ref6","doi-asserted-by":"crossref","first-page":"152","DOI":"10.1128\/MMBR.56.1.152-179.1992","article-title":"Evolution and ecology of influenza A viruses","volume":"56","author":"RG Webster","year":"1992","journal-title":"Microbiol Rev"},{"key":"ref7","doi-asserted-by":"crossref","unstructured":"Wu Y, Tefsen B, Shi Y, Gao GF (2014) Bat-derived influenza-like viruses H17N10 and H18N11. Trends Microbiol.","DOI":"10.1016\/j.tim.2014.01.010"},{"key":"ref8","doi-asserted-by":"crossref","first-page":"893","DOI":"10.1177\/0300985810378651","article-title":"Comparative pathology of select agent influenza a virus infections","volume":"47","author":"T Kuiken","year":"2010","journal-title":"Vet Pathol"},{"key":"ref9","doi-asserted-by":"crossref","first-page":"2628","DOI":"10.1056\/NEJMoa0906089","article-title":"Outbreak of 2009 pandemic influenza A (H1N1) at a New York City school","volume":"361","author":"J Lessler","year":"2009","journal-title":"N Engl J Med"},{"key":"ref10","doi-asserted-by":"crossref","first-page":"393","DOI":"10.1126\/science.279.5349.393","article-title":"Characterization of an avian influenza A (H5N1) virus isolated from a child with a fatal respiratory illness","volume":"279","author":"K Subbarao","year":"1998","journal-title":"Science"},{"key":"ref11","doi-asserted-by":"crossref","first-page":"f2151","DOI":"10.1136\/bmj.f2151","article-title":"H7N9 avian flu infects humans for the first time","volume":"346","author":"J Parry","year":"2013","journal-title":"BMJ"},{"key":"ref12","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1016\/j.jinf.2013.12.014","article-title":"Emergence in China of human disease due to avian influenza A(H10N8)\u2014Cause for concern?","volume":"68","author":"KK To","year":"2014","journal-title":"J Infect"},{"key":"ref13","doi-asserted-by":"crossref","first-page":"9679","DOI":"10.1128\/JVI.73.11.9679-9682.1999","article-title":"Rescue of influenza A virus from recombinant DNA","volume":"73","author":"E Fodor","year":"1999","journal-title":"J Virol"},{"key":"ref14","doi-asserted-by":"crossref","first-page":"9345","DOI":"10.1073\/pnas.96.16.9345","article-title":"Generation of influenza A viruses entirely from cloned cDNAs","volume":"96","author":"G Neumann","year":"1999","journal-title":"Proc Natl Acad Sci U S A"},{"key":"ref15","doi-asserted-by":"crossref","first-page":"4188","DOI":"10.1128\/JVI.70.6.4188-4192.1996","article-title":"A plasmid-based reverse genetics system for influenza A virus","volume":"70","author":"S Pleschka","year":"1996","journal-title":"J Virol"},{"key":"ref16","first-page":"553","article-title":"Antigenic Variation among Avian Influenza a Viruses","volume":"37","author":"HG Pereira","year":"1967","journal-title":"Bulletin of the World Health Organization"},{"key":"ref17","doi-asserted-by":"crossref","first-page":"635","DOI":"10.1016\/j.coviro.2011.07.003","article-title":"Predicting \u2018airborne\u2019 influenza viruses: (trans-) mission impossible?","volume":"1","author":"EM Sorrell","year":"2011","journal-title":"Curr Opin Virol"},{"key":"ref18","doi-asserted-by":"crossref","first-page":"1851","DOI":"10.1128\/JVI.78.4.1851-1857.2004","article-title":"Generation of high-yielding influenza A viruses in African green monkey kidney (Vero) cells by reverse genetics","volume":"78","author":"H Ozaki","year":"2004","journal-title":"J Virol"},{"key":"ref19","doi-asserted-by":"crossref","first-page":"192","DOI":"10.1006\/viro.2002.1742","article-title":"Evaluation of a genetically modified reassortant H5N1 influenza A virus vaccine candidate generated by plasmid-based reverse genetics","volume":"305","author":"K Subbarao","year":"2003","journal-title":"Virology"},{"key":"ref20","doi-asserted-by":"crossref","first-page":"3165","DOI":"10.1016\/S0264-410X(02)00268-2","article-title":"Eight-plasmid system for rapid generation of influenza virus vaccines","volume":"20","author":"E Hoffmann","year":"2002","journal-title":"Vaccine"},{"key":"ref21","doi-asserted-by":"crossref","first-page":"11411","DOI":"10.1073\/pnas.172393399","article-title":"Rescue of influenza B virus from eight plasmids","volume":"99","author":"E Hoffmann","year":"2002","journal-title":"Proc Natl Acad Sci U S A"},{"key":"ref22","doi-asserted-by":"crossref","first-page":"625","DOI":"10.1002\/(SICI)1097-0290(19980620)58:6<625::AID-BIT8>3.0.CO;2-K","article-title":"Plasmid maintenance in Escherichia coli recombinant cultures is dramatically, steadily, and specifically influenced by features of the encoded proteins","volume":"58","author":"JL Corchero","year":"1998","journal-title":"Biotechnol Bioeng"},{"key":"ref23","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1006\/geno.1996.0338","article-title":"Tandem repeats 3\u2032 of the IGHA genes in the human immunoglobulin heavy chain gene cluster","volume":"35","author":"HK Kang","year":"1996","journal-title":"Genomics"},{"key":"ref24","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1016\/0378-1119(94)90857-5","article-title":"Instability of plasmid DNA maintenance caused by transcription of poly(dT)-containing sequences in Escherichia coli","volume":"150","author":"R Kiyama","year":"1994","journal-title":"Gene"},{"key":"ref25","doi-asserted-by":"crossref","first-page":"939","DOI":"10.1016\/0092-8674(82)90456-1","article-title":"Promoter occlusion: transcription through a promoter may inhibit its activity","volume":"29","author":"S Adhya","year":"1982","journal-title":"Cell"},{"key":"ref26","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1016\/S0079-6603(00)66029-4","article-title":"The intrinsically unstable life of DNA triplet repeats associated with human hereditary disorders","volume":"66","author":"RP Bowater","year":"2001","journal-title":"Prog Nucleic Acid Res Mol Biol"},{"key":"ref27","doi-asserted-by":"crossref","first-page":"1399","DOI":"10.1002\/j.1460-2075.1982.tb01329.x","article-title":"Transcription from efficient promoters can interfere with plasmid replication and diminish expression of plasmid specified genes","volume":"1","author":"D Stueber","year":"1982","journal-title":"EMBO J"},{"key":"ref28","doi-asserted-by":"crossref","first-page":"639","DOI":"10.1007\/s004380050858","article-title":"Genetic stability and DNA rearrangements associated with a 2 \u00d7 1.1-Kb perfect palindrome in Escherichia coli","volume":"259","author":"F Malagon","year":"1998","journal-title":"Mol Gen Genet"},{"key":"ref29","doi-asserted-by":"crossref","first-page":"401","DOI":"10.1128\/MMBR.58.3.401-465.1994","article-title":"Biochemistry of homologous recombination in Escherichia coli","volume":"58","author":"SC Kowalczykowski","year":"1994","journal-title":"Microbiol Rev"},{"key":"ref30","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1146\/annurev.biochem.71.083101.133940","article-title":"The bacterial RecA protein and the recombinational DNA repair of stalled replication forks","volume":"71","author":"SL Lusetti","year":"2002","journal-title":"Annu Rev Biochem"},{"key":"ref31","doi-asserted-by":"crossref","first-page":"451","DOI":"10.1073\/pnas.53.2.451","article-title":"Isolation and Characterization of Recombination-Deficient Mutants of Escherichia Coli K12","volume":"53","author":"AJ Clark","year":"1965","journal-title":"Proc Natl Acad Sci U S A"},{"key":"ref32","doi-asserted-by":"crossref","first-page":"1137","DOI":"10.1093\/genetics\/53.6.1137","article-title":"Mutants of Escherichia coli K-12 defective in DNA repair and in genetic recombination","volume":"53","author":"P Howard-Flanders","year":"1966","journal-title":"Genetics"},{"key":"ref33","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1016\/1046-5928(91)90008-7","article-title":"Effect of temperature on the expression of wild-type and thermostable mutants of kanamycin nucleotidyltransferase in Escherichia coli","volume":"2","author":"HH Liao","year":"1991","journal-title":"Protein Expr Purif"},{"key":"ref34","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1016\/0378-1119(93)90766-V","article-title":"Lentivirus envelope sequences and proviral genomes are stabilized in Escherichia coli when cloned in low-copy-number plasmid vectors","volume":"124","author":"TP Cunningham","year":"1993","journal-title":"Gene"},{"key":"ref35","doi-asserted-by":"crossref","DOI":"10.2144\/02325bm03","article-title":"Increased efficiency of cloning large DNA fragments using a lower copy number plasmid","volume":"32","author":"T Feng","year":"2002","journal-title":"Biotechniques"},{"key":"ref36","doi-asserted-by":"crossref","first-page":"378","DOI":"10.1016\/j.jviromet.2011.01.021","article-title":"Reverse genetics plasmid for cloning unstable influenza A virus gene segments","volume":"173","author":"B Zhou","year":"2011","journal-title":"J Virol Methods"},{"key":"ref37","doi-asserted-by":"crossref","first-page":"310","DOI":"10.1006\/viro.1999.0140","article-title":"\u201cAmbisense\u201d approach for the generation of influenza A virus: vRNA and mRNA synthesis from one template","volume":"267","author":"E Hoffmann","year":"2000","journal-title":"Virology"},{"key":"ref38","doi-asserted-by":"crossref","first-page":"2275","DOI":"10.1007\/s007050170002","article-title":"Universal primer set for the full-length amplification of all influenza A viruses","volume":"146","author":"E Hoffmann","year":"2001","journal-title":"Arch Virol"},{"key":"ref39","doi-asserted-by":"crossref","first-page":"603","DOI":"10.1016\/j.jviromet.2013.07.021","article-title":"Improved dual promotor-driven reverse genetics system for influenza viruses","volume":"193","author":"A Mostafa","year":"2013","journal-title":"J Virol Methods"},{"key":"ref40","doi-asserted-by":"crossref","first-page":"e115","DOI":"10.1093\/nar\/gnh110","article-title":"An efficient one-step site-directed and site-saturation mutagenesis protocol","volume":"32","author":"L Zheng","year":"2004","journal-title":"Nucleic Acids Res"},{"key":"ref41","doi-asserted-by":"crossref","first-page":"547","DOI":"10.1128\/JVI.74.1.547-551.2000","article-title":"Plasmid-driven formation of influenza virus-like particles","volume":"74","author":"G Neumann","year":"2000","journal-title":"J Virol"},{"key":"ref42","doi-asserted-by":"crossref","first-page":"1107","DOI":"10.1016\/0092-8674(89)90766-6","article-title":"Amplification, expression, and packaging of foreign gene by influenza virus","volume":"59","author":"W Luytjes","year":"1989","journal-title":"Cell"},{"key":"ref43","doi-asserted-by":"crossref","first-page":"301","DOI":"10.1038\/35060098","article-title":"Influenza virus propagation is impaired by inhibition of the Raf\/MEK\/ERK signalling cascade","volume":"3","author":"S Pleschka","year":"2001","journal-title":"Nature Cell Biology"},{"key":"ref44","doi-asserted-by":"crossref","first-page":"6108","DOI":"10.1073\/pnas.100133697","article-title":"A DNA transfection system for generation of influenza A virus from eight plasmids","volume":"97","author":"E Hoffmann","year":"2000","journal-title":"Proc Natl Acad Sci U S A"},{"key":"ref45","doi-asserted-by":"crossref","first-page":"2122","DOI":"10.1128\/JVI.01668-09","article-title":"The NS segment of an H5N1 highly pathogenic avian influenza virus (HPAIV) is sufficient to alter replication efficiency, cell tropism, and host range of an H7N1 HPAIV","volume":"84","author":"W Ma","year":"2010","journal-title":"J Virol"},{"key":"ref46","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1084\/jem.75.1.49","article-title":"The Quantitative Determination of Influenza Virus and Antibodies by Means of Red Cell Agglutination","volume":"75","author":"GK Hirst","year":"1942","journal-title":"J Exp Med"},{"key":"ref47","doi-asserted-by":"crossref","first-page":"2189","DOI":"10.1007\/s00705-008-0243-2","article-title":"Antigenic structure of the hemagglutinin of H9N2 influenza viruses","volume":"153","author":"M Okamatsu","year":"2008","journal-title":"Archives of Virology"},{"key":"ref48","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1016\/0378-1119(94)90235-6","article-title":"Positive-selection vectors using the F plasmid ccdB killer gene","volume":"148","author":"P Bernard","year":"1994","journal-title":"Gene"},{"key":"ref49","doi-asserted-by":"crossref","first-page":"4786","DOI":"10.1128\/JVI.03551-12","article-title":"A Single Amino Acid at the Hemagglutinin Cleavage Site Contributes to the Pathogenicity but Not the Transmission of Egyptian Highly Pathogenic H5N1 Influenza Virus in Chickens","volume":"87","author":"SW Yoon","year":"2013","journal-title":"J Virol"},{"key":"ref50","doi-asserted-by":"crossref","first-page":"1414","DOI":"10.1371\/journal.ppat.0030141","article-title":"A single mutation in the PB1-F2 of H5N1 (HK\/97) and 1918 influenza A viruses contributes to increased virulence","volume":"3","author":"GM Conenello","year":"2007","journal-title":"PLoS Pathog"},{"key":"ref51","doi-asserted-by":"crossref","first-page":"1840","DOI":"10.1126\/science.1062882","article-title":"Molecular basis for high virulence of Hong Kong H5N1 influenza A viruses","volume":"293","author":"M Hatta","year":"2001","journal-title":"Science"},{"key":"ref52","doi-asserted-by":"crossref","first-page":"2943","DOI":"10.1016\/j.vaccine.2004.08.054","article-title":"Generation of influenza vaccine viruses on Vero cells by reverse genetics: an H5N1 candidate vaccine strain produced under a quality system","volume":"23","author":"C Nicolson","year":"2005","journal-title":"Vaccine"},{"key":"ref53","doi-asserted-by":"crossref","first-page":"1524","DOI":"10.1038\/sj.gt.3302807","article-title":"Overcoming promoter competition in packaging cells improves production of self-inactivating retroviral vectors","volume":"13","author":"A Schambach","year":"2006","journal-title":"Gene Ther"},{"key":"ref54","doi-asserted-by":"crossref","first-page":"379","DOI":"10.1128\/MCB.7.1.379","article-title":"Analysis of mutation in human cells by using an Epstein-Barr virus shuttle system","volume":"7","author":"RB DuBridge","year":"1987","journal-title":"Mol Cell Biol"},{"key":"ref55","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1007\/s13205-012-0070-8","article-title":"Remarkable stability of an instability-prone lentiviral vector plasmid in Escherichia coli Stbl3","volume":"3","author":"FA Al-Allaf","year":"2013","journal-title":"3 Biotech"},{"key":"ref56","doi-asserted-by":"crossref","first-page":"6542","DOI":"10.1128\/JVI.00641-13","article-title":"Chimeric hemagglutinin influenza virus vaccine constructs elicit broadly protective stalk-specific antibodies","volume":"87","author":"F Krammer","year":"2013","journal-title":"J Virol"},{"key":"ref57","doi-asserted-by":"crossref","first-page":"8008","DOI":"10.1016\/j.vaccine.2010.09.006","article-title":"Improved haemagglutinin antigen content in H5N1 candidate vaccine viruses with chimeric haemagglutinin molecules","volume":"28","author":"R Harvey","year":"2010","journal-title":"Vaccine"},{"key":"ref58","doi-asserted-by":"crossref","first-page":"6086","DOI":"10.1128\/JVI.00096-11","article-title":"Improved antigen yield in pandemic H1N1 (2009) candidate vaccine viruses with chimeric hemagglutinin molecules","volume":"85","author":"R Harvey","year":"2011","journal-title":"J Virol"},{"key":"ref59","doi-asserted-by":"crossref","first-page":"459","DOI":"10.1016\/0022-2836(69)90288-5","article-title":"A complementation analysis of the restriction and modification of DNA in Escherichia coli","volume":"41","author":"HW Boyer","year":"1969","journal-title":"J Mol Biol"},{"key":"ref60","doi-asserted-by":"crossref","first-page":"6313","DOI":"10.1073\/pnas.79.20.6313","article-title":"Control of ColE1 DNA replication: the rop gene product negatively affects transcription from the replication primer promoter","volume":"79","author":"G Cesareni","year":"1982","journal-title":"Proc Natl Acad Sci U S A"},{"key":"ref61","doi-asserted-by":"crossref","DOI":"10.1186\/1472-6750-5-19","article-title":"Viral promoters can initiate expression of toxin genes introduced into Escherichia coli","volume":"5","author":"A Lewin","year":"2005","journal-title":"Bmc Biotechnology"},{"key":"ref62","doi-asserted-by":"crossref","first-page":"e139","DOI":"10.1093\/nar\/gkn646","article-title":"Rapid and reliable universal cloning of influenza A virus genes by target-primed plasmid amplification","volume":"36","author":"J Stech","year":"2008","journal-title":"Nucleic Acids Res"},{"key":"ref63","doi-asserted-by":"crossref","first-page":"e5553","DOI":"10.1371\/journal.pone.0005553","article-title":"Golden gate shuffling: a one-pot DNA shuffling method based on type IIs restriction enzymes","volume":"4","author":"C Engler","year":"2009","journal-title":"PLoS One"},{"key":"ref64","doi-asserted-by":"crossref","first-page":"e3647","DOI":"10.1371\/journal.pone.0003647","article-title":"A one pot, one step, precision cloning method with high throughput capability","volume":"3","author":"C Engler","year":"2008","journal-title":"PLoS One"}],"container-title":["PLOS ONE"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/dx.plos.org\/10.1371\/journal.pone.0116917","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,4,27]],"date-time":"2022-04-27T12:39:57Z","timestamp":1651063197000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0116917"}},"subtitle":[],"editor":[{"given":"Florian","family":"Krammer","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2015,1,23]]},"references-count":64,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2015,1,23]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0116917","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2015,1,23]]}}}