Scientific papers // The enzymes genes cloning and study // Cloning and Study of New DNA Methyltransferase M.FatI Modifying Cytosine in a Recognition Site CATG
Cloning and Study of New DNA Methyltransferase M.FatI Modifying Cytosine in a Recognition Site CATG
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Dedkov V.S., Gonchar D.A., Abdurashitov M.A., Udalyeva S.G., Urumceva L.A., Chernukhin V.A., Mutylo G.V., Degtyarev S.Kh
Research Journal of Pharmaceutical, Biological and Chemical Sciences, vol 6 (6), 2015 P. 1341-1348
A fragment of Flavobacterium aquatile NL3 DNA carrying the gene of DNA methyltransferase M.FatI was cloned in pUC19 plasmid. DNA was sequenced and M.FatI gene was analyzed. A recombinant strain Esherichia coli was grown up and the enzyme was purified. M.FatI specificity was determined by a blocking of some restriction endonucleases and computer modeling. It’s well known that M.NlaIII produces 5’-C(m6A)TG- 3’, whereas FatI MTase modifies the cytosine residue with formation 5’-(m5C)ATG-3’. The sensitivity of restriction endonucleases to FatI-methylation has been studied. Keywords: gene cloning, enzyme isolation, bacterial DNA methyltransferase, enzyme specificity, restriction endonuclease, methylation sensitivity