Jf. Glickman et No. Reich, BACULOVIRUS-MEDIATED HIGH-LEVEL EXPRESSION OF A MAMMALIAN DNA METHYLTRANSFERASE, Biochemical and biophysical research communications, 204(3), 1994, pp. 1003-1008
The murine C-5 cytosine DNA methyltransferase (MTase, E.C.2.1.1.37) co
ntaining a hexahistidine affinity leader peptide has been expressed at
levels which are at least 50-fold higher than previously reported. Th
e recombinant enzyme has activity levels similar to the wild-type enzy
me. The recombinant polypeptide binds to and elutes from a nickel affi
nity resin (IMAC resin). No dramatic differences in post-translational
modification between the wild-type and recombinant enzyme were observ
ed. The recombinant system will be useful in performing site-directed
mutagenesis and will facilitate enzymological and biological investiga
tions of this enzyme. (C) 1994 Academic Press, Inc.