Generation of recombinant influenza virus using baculovirus delivery vector.
A recombinant baculovirus vector containing mammalian cell-active promoters and transcription terminators was used to deliver a mutated influenza NS gene into Vero cells. In addition to the influenza NS gene, the baculovirus contained a reporter gene expression cassette (Green fluorescent protein, GFP), allowing to monitor the Vero cell transduction efficiency. More than 90% of Vero cells were expressing GFP 24-48 h post transduction. After infecting baculovirus transduced cells with influenza helper virus, progeny of attenuated influenza virus carrying the recombinant NS gene could be selected. Baculovirus delivery was highly reproducible and efficient in Vero cells. This new method for influenza gene delivery could contribute to influenza virus research and vaccine development.
Poomputsa
Kanokwan
K
School of Bioresources and Technology, King Mongkut's University of Technology Thonburi, 83 Moo 8 Thakham, Bangkhuntien, 10150, Bangkok, Thailand.
Kittel
Christian
C
Egorov
Andrej
A
Ernst
Wolfgang
W
Grabherr
Reingard
R
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D002522:Chlorocebus aethiops; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D009980:Influenza A virus / Q000235:genetics*; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic; D014161:Transduction, Genetic; D014709:Vero Cells; D017361:Viral Nonstructural Proteins / Q000235:genetics* / Q000378:metabolism
C085820:INS1 protein, influenza virus; D008164:Luminescent Proteins; D017361:Viral Nonstructural Proteins; D049452:Green Fluorescent Proteins