Biochemical method for mapping mutational alterations in DNA with S1 nuclease: the location of deletions and temperature sensitive mutations in simian virus 40

T. E. Shenk, C. Rhodes, P. W.J. Rigby, P. Berg

Research output: Contribution to journalArticle

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Abstract

S1 nuclease (EC 3.1.4.X), a single strand specific nuclease, can be used to accurately map the location of mutational alterations in simian virus 40 (SV40) DNA. Deletions of between 32 and 190 base pairs, which are at or below the limit of detectability by conventional electron microscopic analysis of heteroduplex DNAs, have been located in this way. To map a deletion, a mixture of unit length, linear DNA, prepared from the SV40 deletion mutant and its wild type parent, are denatured and reannealed to form heteroduplexes. S1 nuclease can cut such heteroduplexes at the nonbase paired region to produce fragments whose lengths correspond to the position of the deletion. Similarly specific fragments are produced when S1 nuclease cleaves a heteroduplex formed from the DNAs of SV40 temperature sensitive mutants and either their revertants or wild type parents. Thus, the positions of the nonhomology between these DNAs can be determined.

Original languageEnglish (US)
Pages (from-to)989-993
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume72
Issue number3
DOIs
StatePublished - 1975

All Science Journal Classification (ASJC) codes

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