title: A one-plasmid system to generate influenza virus in cultured chicken cells for potential use in influenza vaccine.
abstract: Influenza virus has a set of ribonucleoproteins (RNPs) consisting of viral RNAs, influenza virus polymerase subunits, and nucleoprotein. Intracellular reconstitution of the whole set of RNPs via plasmid transfection results in the generation of influenza virus. By the use of reverse genetics and dual promoters, we constructed a 23.6-kb eight-unit plasmid that contains all the required constituents to generate influenza virus in chicken cells. Our "one-plasmid" system generated higher titers of influenza virus in chicken cells than the "eight-plasmid" system, enabling a simpler approach for generating vaccine seeds. Our study identified plasmid size as a potential limiting factor affecting transfection efficiency and hence the influenza viral yield from chicken cells.
authors: Center for Infectious Diseases and Vaccinology, the Biodesign Institute, Arizona State University, Tempe, 85287-5401, USA.
authors: Xiangmin
authors: X
authors: Zhang
authors: Wei
authors: W
authors: Kong
authors: Shamaila
authors: S
authors: Ashraf
authors: Roy
authors: R
authors: Curtiss
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D002645:Chickens; D008828:Microbiological Techniques; D009975:Orthomyxoviridae; D010957:Plasmids / Q000235:genetics*; D011401:Promoter Regions, Genetic; D014162:Transfection; D014773:Virology / Q000379:methods*; D014779:Virus Replication