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Nucleic Acids Research, 1987, Vol. 15, No. 17 7091-7102
© 1987

Articles

Cleavage of methylated CCCGGG sequences containing either N4-methylcytosine or 5-methytcytosine with Mspl, Hpall, Smal, Xmal and Cfr9I restriction endonudeases

V. Butkus, L. Petrauskiene, Z. Maneliene, S. Klimasauskas, V. Laucys and A. Janulaitis

ESP Fennentas, All Union Research Institute of Applied Enzymology Fermentu 8, Vilnius 232028, Lithuanian SSR, USSR

Received June 5, 1987. Revised August 5, 1987. Accepted August 5, 1987.

The cleavage specificity of R.Cfr9I was determined to be CCGGG whereas the methylation specificity of H.Cfr9I was C4mCCGGG. The action of HspI, HpaII, SmaI, XmaI and Cfr9l restriction endonudeases on an unmethylated parent d(GGACCCGGGTCC) dodecanucleotide duplex and a set of oligo-nuoleotide duplexes, containing all possible substitutions of either 4mC or 5mC for C in the CCCGGG sequence, was investigated. It was found that 4mC methylation. in contrast to 5mC, renders the CCCGGG site resistant to practically all the investigated endonuoleases. The cleavage of methylated substrates with restriction endonucleases is discussed.


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