Tarasova M.V.,Novosibirsk State University |
Kuznetsov V.V.,Federal State Institution of Science |
Netesova N.A.,Federal State Institution of Science |
Gonchar D.A.,SibEnzyme Scientific Production Association |
Degtyarev S.K.,SibEnzyme Scientific Production Association
Moscow University Biological Sciences Bulletin | Year: 2011
DNA methyltransferases genes of the BspACI restriction-modification system from Bacillus psychrodurans AC have been cloned in E. coli cells. Analysis of amino acid sequences of the proteins showed that both of these genes belong to C5 DNA methyltransferases. Gene M1. BspACI has been subcloned in pJW2 vector. A high-purity recombinant enzyme has been obtained using chromatography on different carriers. It has been shown that M1. BspACI modifies the first cytosine residue in the sequence 5′-CCGC-3′. Kinetic parameters of DNA methylation by the enzyme have been determined. Catalytic constant appears to be 0.095 ± 0.002 min-1. Km phage is λ DNA-0.053 ± 0.007 μM, and KmSAM is 5.1 ± 0.3 μM. © 2011 Allerton Press, Inc.