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Biochemical Procedure for Production of Small Deletions in Simian Virus 40 DNA

A simple biochemical procedure for producing small deletions (15 to 50 base pairs) at virtually any location in simian virus 40 DNA has been developed. The steps involved are: cleavage of the closed-circular DNA to produce a linear structure followed by 5′-exonuclease digestion to expose a short sin...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1975-04, Vol.72 (4), p.1392-1396
Main Authors: Carbon, John, Shenk, Thomas E., Berg, Paul
Format: Article
Language:English
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Summary:A simple biochemical procedure for producing small deletions (15 to 50 base pairs) at virtually any location in simian virus 40 DNA has been developed. The steps involved are: cleavage of the closed-circular DNA to produce a linear structure followed by 5′-exonuclease digestion to expose a short single-stranded segment at each 3′end of the molecule. Mutants containing deletions at the site of the cleavage are obtained by infecting permissive monkey kidney cells with the exonuclease-treated DNA in the presence or absence of a helper DNA (depending upon whether or not the site of cleavage and therefore the deletion occurred in a gene required for vegetative multiplication). In this paper viable mutants with deletions at the HpaII endonuclease cleavage site (0.735 map position) and defective trans-complementable mutants with deletions at the EcoRI endonuclease cleavage site (0/1.0 map position) were isolated.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.72.4.1392