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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
6
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pubmed:dateCreated |
1996-7-23
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pubmed:abstractText |
A reverse genetics system for negative-strand RNA viruses was first successfully developed for influenza viruses. This technology involved the transfection of in vitro-reconstituted ribonucleoprotein (RNP) complexes into influenza virus-infected cells. We have now developed a method that allows intracellular reconstitution of RNP complexes from plasmid-based expression vectors. Expression of a viral RNA-like transcript is achieved from a plasmid containing a truncated human polymerase I (polI) promoter and a ribozyme sequence that generates the desired 3' end by autocatalytic cleavage. The polI-driven plasmid is cotransfected into human 293 cells with polII-responsive plasmids that express the viral PB1, PB2, PA, and NP proteins. This exclusively plasmid-driven system results in the efficient transcription and replication of the viral RNA-like reporter and allows the study of cis- and trans-acting signals involved in the transcription and replication of influenza virus RNAs. Using this system, we have also been able to rescue a synthetic neuraminidase gene into a recombinant influenza virus. This method represents a convenient alternative to the previously established RNP transfection system.
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-1607855,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-1629705,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-1727600,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-2011192,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-2016777,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-2033659,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-2052614,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-2214032,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-2339122,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-2598262,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-2650474,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-2813075,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-2983190,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-3422449,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-561860,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-7483820,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-7483828,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-7507179,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-7535862,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-7667300,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-7753828,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-7815505,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-7925265,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-7966605,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-8009859,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-8046417,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-8107244,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-8166749,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-8230444,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-8257115,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-8367275,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-8383187,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-8524804,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-8846771,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8648766-9049376
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Jun
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pubmed:issn |
0022-538X
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
70
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
4188-92
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:8648766-Base Sequence,
pubmed-meshheading:8648766-Humans,
pubmed-meshheading:8648766-Influenza A virus,
pubmed-meshheading:8648766-Molecular Sequence Data,
pubmed-meshheading:8648766-Plasmids,
pubmed-meshheading:8648766-RNA, Viral,
pubmed-meshheading:8648766-Ribonucleoproteins,
pubmed-meshheading:8648766-Transcription, Genetic,
pubmed-meshheading:8648766-Transfection
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pubmed:year |
1996
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pubmed:articleTitle |
A plasmid-based reverse genetics system for influenza A virus.
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pubmed:affiliation |
Department of Microbiology, Mount Sinai School of Medicine, New York 10029, USA.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
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