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Nucleic Acids Research, 1985, Vol. 13, No. 12 4285-4298
© 1985

Articles

Abasic sites from cytosine as termination signals for DNA synthesis

Daphna Sagher and Bernard Strauss*

Department of Molecular Genetics and Cell Biology, University of Chicago Chicago, IL 60631, USA

*To whom correspondence should be addressed

Received April 15, 1985. Accepted May 29, 1985.

DNA with abasic sites has been prepared by deamination of cytosine followed by treatment of the product with uracil N-glycosylase. Termination in vitro on such templates does not occur until treatment with uracil N-glycosylase. DNA terminated one base before abasic sites created from C's has been used as a template in "second stage" reactions. With enzymes devoid or deficient in 3'>5' exonuclease activity purines, particularly adenine, are preferentially added opposite the putative abasic site. 2-Aminopurine behaves more like adenine than like guanine in these experiments. Polyraerase ß preferentially incorporates A opposite abasic sites produced from T, and G opposite abasic sites produced from C. We have eliminated an obvious artefact (e.g. strand switching) which might account for this observation


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