title: Rapid regeneration of virus from cells infected with a retroviral vector.
abstract: Recombinant retroviral vectors usually encode the genes of interest in place of the viral structural genes, which must be provided in trans. These viruses are therefore defective for replication: infected cells cannot produce progeny virus. However, in some cases it may be desirable to generate virus from an infected cell clone displaying a phenotype of interest. We describe a rapid method for producing virus, which involves fusing the infected cells to fresh packaging cells. Stable producer lines are generated after fusion by co-selecting for the Ecogpt+ marker in the packaging cells and the G418 resistance (neor) marker in the infected cells. We have used this method to develop cell lines that produce retroviruses encoding a Leu 61-activated c-Ha-ras oncogene as well as a neor gene. The viruses confer oncogenic transformation on 95%-100% of infected target cells as assayed by altered morphology, focus formation and soft agar growth.
authors.affiliation: Division of Cellular and Molecular Biology, Dana-Farber Cancer Institute, Boston, MA 02115.
authors.forename: M J
authors.initials: MJ
authors.lastname: Corbley
authors.forename: V
authors.initials: V
authors.lastname: Cherington
authors.forename: L A
authors.initials: LA
authors.lastname: Feig
authors.forename: G M
authors.initials: GM
authors.lastname: Cooper
authors.forename: T M
authors.initials: TM
authors.lastname: Roberts
mesh_terms: D000818:Animals; D002460:Cell Line; D004261:DNA Replication / Q000235:genetics*; D004279:DNA, Viral / Q000096:biosynthesis*; D005822:Genetic Vectors; D015689:Oncogene Protein p21(ras) / Q000032:analysis; D012190:Retroviridae / Q000502:physiology*; D014779:Virus Replication / Q000235:genetics*
chemical_list: D004279:DNA, Viral; D015689:Oncogene Protein p21(ras)