BACULOVIRUS-MEDIATED HIGH-LEVEL EXPRESSION OF A MAMMALIAN DNA METHYLTRANSFERASE

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.