TY - JOUR
T1 - Biochemical method for mapping mutational alterations in DNA with S1 nuclease
T2 - the location of deletions and temperature sensitive mutations in simian virus 40
AU - Shenk, T. E.
AU - Rhodes, C.
AU - Rigby, P. W.J.
AU - Berg, P.
PY - 1975
Y1 - 1975
N2 - 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.
AB - 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.
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U2 - 10.1073/pnas.72.3.989
DO - 10.1073/pnas.72.3.989
M3 - Article
C2 - 165498
AN - SCOPUS:1642591341
SN - 0027-8424
VL - 72
SP - 989
EP - 993
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 3
ER -