In vitro reconstitution of active influenza virus ribonucleoprotein complexes using viral proteins purified from infected cells.
A procedure to obtain RNA-free preparations containing the nucleoprotein (NP) and polymerase (P) proteins from influenza virus-infected cell extracts has been developed. The influenza virus P proteins present in these preparations copied small synthetic RNA molecules derived from plasmid sequences. In addition, RNA molecules encapsidated with the NP and P proteins were amplified and packaged into virus particles when transfected into influenza virus-infected cells. Thus, the preparations of NP and P proteins display features similar to those isolated from purified influenza virions, and represent an alternative for the preparation of active influenza virus P protein.
Martín
J
J
Instituto de Salud Carlos III, Centro Nacional de Microbiología, Madrid, Spain.
Albo
C
C
Ortín
J
J
Melero
J A
JA
Portela
A
A
D000818:Animals; D002478:Cells, Cultured; D015500:Chloramphenicol O-Acetyltransferase / Q000032:analysis; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D009698:Nucleoproteins / Q000378:metabolism*; D009975:Orthomyxoviridae / Q000378:metabolism*; D012367:RNA, Viral / Q000378:metabolism*; D012261:Ribonucleoproteins / Q000378:metabolism*; D014764:Viral Proteins / Q000302:isolation & purification / Q000378:metabolism*
D009698:Nucleoproteins; D012367:RNA, Viral; D012261:Ribonucleoproteins; D014764:Viral Proteins; D015500:Chloramphenicol O-Acetyltransferase; D012321:DNA-Directed RNA Polymerases