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PMID: 6270359 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Construction and isolation of a transmissible retrovirus containing the src gene of Harvey murine sarcoma virus and the thymidine kinase gene of herpes simplex virus type 1.

Journal of virology ·Vol. 39 ·No. 3 ·1981-09-00 ·Pages 935-44

Wei CM, Gibson M, Spear PG, Scolnick EM

Abstract

We constructed lambda recombinants containing the Harvey murine sarcoma virus genome and the thymidine kinase (tk) gene of herpes simplex virus type 1 linked to each other. The tk gene was located in a position downstream from both the long terminal repeat and the src gene of Harvey murine sarcoma virus. The DNAs of the lambda recombinants were used to transfect NIH3T3 mouse fibroblasts in order to obtain Harvey murine sarcoma virus DNA-induced foci of transformed cells. The transformed foci were superinfected with a helper-independent retrovirus, and new individual retrovirus were isolated from the superinfected foci. The new viruses could induce focus formation on NIH3T3 cells and could convert NIH3T3(TK-) cells into TK+ cells by carrying the herpes simplex virus type 1 tk gene into the TK- cells. From virus-infected cells, we isolated nonproducer foci on NIH3T3 cells and TK+ transformants on NIH3T3(TK-) cells containing one such new viral genome coding for the dual properties. The new retroviral sequence in the nonproducer cells could be rescued into virus particles at high titers by superinfection with a helper-independent retrovirus. A hybridization analysis indicated that the recombinant virus contained both the Harvey murine sarcoma virus src sequence and the tk gene sequence in a single RNA species approximately 4.9 kilobases long. We concluded that retroviruses can be used as true vectors for genes other than genes that lead to oncogenesis.

MeSH Terms
Animals Cell Line Cell Transformation, Viral Cloning, Molecular DNA, Recombinant Genes, Viral Genetic Vectors Mice Retroviridae/genetics,isolation & purification Sarcoma Viruses, Murine/genetics Simplexvirus/genetics Thymidine Kinase/genetics
Chemicals
DNA, Recombinant Thymidine Kinase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Wei C M
Gibson M
Spear P G
Scolnick E M
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48 references, click to expand
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.39.3.935-944.1981
Published
1981-09-00
Pages
935-44
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC171327
Subset
IM
Grants
NCI NIH HHS · CA 21776 · United States
External Links
PubMed source DOI: 10.1128/JVI.39.3.935-944.1981 PMC Full Text View journal details
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