title: Infectious alphavirus production from a simple plasmid transfection+.
abstract: We have developed a new method for producing infectious double subgenomic alphaviruses from plasmids transfected into mammalian cells. A double subgenomic Sindbis virus (TE3'2J) was transcribed from a cytomegalovirus PolII promoter, which results in the production of infectious virus. Transfection of as little as 125 ng of plasmid is able to produce 1 × 10(8) plaque forming units/ml (PFU/ml) of infectious virus 48 hours post-transfection. This system represents a more efficient method for producing recombinant Sindbis viruses.
authors.affiliation: Department of Microbiology, Immunology, and Pathology, 1682 Campus Delivery, Colorado State University, Fort Collins, CO 80523, USA.
authors.forename: J Jordan
authors.initials: JJ
authors.lastname: Steel
authors.forename: Brittney R
authors.initials: BR
authors.lastname: Henderson
authors.forename: Siddhi B C
authors.initials: SB
authors.lastname: Lama
authors.forename: Ken E
authors.initials: KE
authors.lastname: Olson
authors.forename: Brian J
authors.initials: BJ
authors.lastname: Geiss
mesh_terms: D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D006224:Cricetinae; D003587:Cytomegalovirus / Q000235:genetics*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012845:Sindbis Virus / Q000235:genetics* / Q000254:growth & development*; D014162:Transfection; D014776:Virus Cultivation / Q000379:methods