Nucleic Acids Research, 1981, Vol. 9, No. 12 2643-2658
© 1981
ENZYMOLOGY |
Sequence specific cleavage of DNA by micrococcal nuclease
Institut für Physiologische Chemie, Physikalische Biochemie und Zellbiologie der Universität München Goethestrasse 33, 8000 München 2, GFR
Received April 23, 1981. Micrococcal nuclease is shown to cleave DNA under conditions of partial digestion in a specific manner. Sequences of the type 5'CATA and 5'CTA are attacked preferentially, followed by exonucleoTytic degradation at the newly generated DNA termini. GC-rich flanking sequences further increase the probability of initial attack. Unexpectedly, long stretches containing only A and T are spared by the nuclease. These results, which were obtained with mouse satellite DNA and two fragments from the plasmid pBR322, do not support the previous contention that it is the regions of high AT-content which are initially cleaved by micrococcal nuclease. This specificity of micrococcal nuclease complicates its use in experiments intended to monitor the nucleoprotein structure of a DNA sequence in chromatin.
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