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Transfection of Fibroblasts by Cloned Abelson Murine Leukemia Virus DNA and Recovery of Transmissible Virus by Recombination with Helper Virus

Goff, Stephen P. and Tabin, Clifford J. and Wang, Jean Yin-Jen and Weinberg, Robert and Baltimore, David (1982) Transfection of Fibroblasts by Cloned Abelson Murine Leukemia Virus DNA and Recovery of Transmissible Virus by Recombination with Helper Virus. Journal of Virology, 41 (1). pp. 271-285. ISSN 0022-538X. PMCID PMC256749. https://resolver.caltech.edu/CaltechAUTHORS:20120717-091825058

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Abstract

A cloned, permuted DNA copy of the Abelson murine leukemia virus (A-MuLV) genome was capable of eliciting the morphological transformation of NIH/3T3 fibroblasts when applied to cells in a calcium phosphate precipitate. The efficiency of the process was extremely low, yielding approximately one transformant per microgram of DNA under conditions which give 104 transfectants per microgram of other DNAs (e.g., Moloney sarcoma virus proviral DNA). The DNA was able to induce foci, even though the 3′ end of the genome was not present. The transforming gene was thus localized to the 5′ portion of the genome. The transformed cells all produced viral RNA and the virus-specific P90 protein. Transmissible virus could be rescued from these cells at very low frequencies by superinfection with helper virus; the rescued A-MuLV virus had variable 3′ ends apparently derived by recombination with the helper. Dimerization of the permuted A-MuLV cloned genome to reconstruct a complete provirus did not improve transformation efficiency. Virus could be rescued from these transformants, however, at a high efficiency. Cotransfection of the permuted A-MuLV DNA with proviral M-MuLV DNA yielded a significant increase in the efficiency of transformation and cotransfection of dimeric A-MuLV and proviral M-MuLV resulted in a high-efficiency transformation yielding several thousand more transformants per microgram than A-MuLV DNA alone. We propose that helper virus efficiently rescues A-MuLV from transiently transfected cells which would not otherwise have grown into foci. We hypothesize that multiple copies of A-MuLV DNA introduced into cells by transfection are toxic to cells. In support of this hypothesis, we have shown that A-MuLV DNA sequences can inhibit the stable transformation of cells by other selectable DNAs.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://jvi.asm.org/content/41/1/271.abstractPublisherArticle
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC256749/PubMed CentralArticle
ORCID:
AuthorORCID
Baltimore, David0000-0001-8723-8190
Additional Information:© 1982 American Society for Microbiology. Received 17 August 1981. Accepted 6 October 1981. We thank Ann Gifford for excellent technical assistance. This work was supported by Public Health Service grant CA-17537 (to R.W.), program project grant CA-26717 (to D. B.), and grant CA-14051 (core grant to S. E. Luria), all from the National Cancer Institute. J.Y.-J.W. is a postdoctoral fellow of the Jane Coffin Childs Memorial Fund for Medical Research. D.B. is an American Cancer Society Research Professor.
Funders:
Funding AgencyGrant Number
NIHCA-17537
NIHCA-26717
NIHCA-14051
Jane Coffin Childs Memorial Fund for Medical ResearchUNSPECIFIED
National Cancer InstituteUNSPECIFIED
Issue or Number:1
PubMed Central ID:PMC256749
Record Number:CaltechAUTHORS:20120717-091825058
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20120717-091825058
Official Citation:Transfection of Fibroblasts by Cloned Abelson Murine Leukemia Virus DNA and Recovery of Transmissible Virus by Recombination with Helper Virus Stephen P. Goff, Clifford J. Tabin, Jean Yin-Jen Wang, Robert Weinberg, and David Baltimore J. Virol. January 1982 41:271-285
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:32505
Collection:CaltechAUTHORS
Deposited By: Ruth Sustaita
Deposited On:17 Jul 2012 16:54
Last Modified:03 Oct 2019 04:01

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