Characteristics of an Sv40-Plasmid Recombinant and Its Movement into and out of the Genome of a Murine Cell

Hanahan, D., Lane, D., Lipsich, L., Wigler, M. H., Botchan, M. (August 1980) Characteristics of an Sv40-Plasmid Recombinant and Its Movement into and out of the Genome of a Murine Cell. Cell, 21 (1). pp. 127-139. ISSN 0092-8674

DOI: 10.1016/0092-8674(80)90120-8


A bacterial plasmid carrying the early region of SV40 (pOT) has been stably established in high molecular weight (hmw) DNA of mouse L cells by selection for the herpes virus thymidine kinase (tk) gene. DNA blotting has demonstrated that most cell lines contain multiple discrete copies of pOT, generally with an intact SV40 early region. No free copies of pOT have been detected. Both pOT and tk sequences may be amplified up to 20-200 copies of the SV40 early region. In contrast to the uniform staining pattern normally observed in SV40-transformed lines, indirect immunofluorescence using antiserum to the SV40 T antigen has demonstrated that the expression of the early region is heterogeneous in these cell lines. This fraction expressing T is characteristic of a given cell line, and varies from 0 to 99% positive. Several pOT cell lines have been fused to simian cells, and replicating low molecular weight DNAs were isolated from the heterokaryons. Transformation of E. coli with this DNA demonstrates that pOT can be rescued from hmw DNA in L cells and reestablished as a plasmid in E. coli. Excision is generally precise when pOT is introduced to the murine cells as supercoiled molecule, and imprecise when pOT is introduced in linear form.

Item Type: Paper
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function > gene transfer
organism description > animal > mammal > rodent > mouse
organism description > virus > viral transfection
organism description > virus
CSHL Authors:
Communities: CSHL labs > Wigler lab
Depositing User: CSHL Librarian
Date: August 1980
Date Deposited: 12 Apr 2012 19:12
Last Modified: 17 Oct 2016 16:06
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