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Nucleic Acids Research, 1990, Vol. 18, No. 8 2159-2162
© 1990

MOLECULAR BIOLOGY

Some restriction endonucleases tolerate single mismatches of the pyrimidine·purine type

M. Petranovic1,2,*, D. Petranovic2, C. Dohet1, P. Brooks1 and M. Radman1

1Institut Jacques Monod T43, 2 place Jussieu, 75251 Paris Cedex 05, France 2Ruder Boskovic Institute PO Box 1016, 41001 Zagreb, Yugoslavia

* To whom correspondence should be addressed

Received October 31, 1989. Revised February 5, 1990. Accepted February 5, 1990.

DNAs from phage mutants M13mp18 and M13mp18/MP-1 were used to construct two closed circular heteroduplexes. One of them carried the sequence 5'-CCTGGG-3' 3'-GGGCCC-5' with a T·G mismatch at the position 6248. The other carried the sequence 5'-CCCGGG-' 3'-GGACCC-5' with a C·A mismatch at the same position. Heteroduplexes were exposed to 7 restriction endonucleases having recognition sites within the sequence 5'-CCCGGG-3' 3'-GGGCCC-5' and to 1 restriction endonuclease having a recognition site within the sequence 5'-CCTGGG-3' 3-GGACCC-5'. All tested enzymes cleaved at least one mismatch-containing sequence although with reduced efficiency. Smal and Xmal tolerated both mismatch-containing sequences. Aval, Hpall, Mspl, Neil and Nspili were able to tolerate only the T·G containing sequence, while BstNl was able to tolerate only the C·A containing sequence. It Is inferred that the tolerance displayed by Smal and Xmal depends on the presence of either the original purines or the original pyrimidines in mismatches of both the T·G and C·A type and that all other tested enzymes require the presence of the original purines in mismaches of both types.


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