Open AccessCleavage of methylated CCCGGG sequences containing either N4-methylcytosine or 5-methytcytosine with Mspl, Hpall, Smal, Xmal and Cfr9I restriction endonudeases
Author(s) -
V. Butkus,
L. Petrauskienė,
Z. Manelienė,
Saulius Klimašauskas,
V. Laučys,
A. Janulaitis
Publication year - 1987
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/15.17.7091
Subject(s) - hpaii , 5 methylcytosine , biology , restriction enzyme , methylation , cleavage (geology) , microbiology and biotechnology , dna methylation , dna , genetics , gene , gene expression , paleontology , fracture (geology)
The cleavage specificity of R.Cfr9I was determined to be C decreases CCGGG whereas the methylation specificity of M.Cfr9I was C4mCCGGG. The action of MspI, HpaII, SmaI, XmaI and Cfr9I restriction endonucleases on an unmethylated parent d(GGACCCGGGTCC) dodecanucleotide duplex and a set of oligonucleotide 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 endonucleases. The cleavage of methylated substrates with restriction endonucleases is discussed.