Glickman J F, Reich N O
Department of Chemistry, University of California, Santa Barbara 93106.
Biochem Biophys Res Commun. 1994 Nov 15;204(3):1003-8. doi: 10.1006/bbrc.1994.2562.
The murine C-5 cytosine DNA methyltransferase (MTase, E.C.2.1.1.37) containing a hexahistidine affinity leader peptide has been expressed at levels which are at least 50-fold higher than previously reported. The recombinant enzyme has activity levels similar to the wild-type enzyme. The recombinant polypeptide binds to and elutes from a nickel affinity resin (IMAC resin). No dramatic differences in post-translational modification between the wild-type and recombinant enzyme were observed. The recombinant system will be useful in performing site-directed mutagenesis and will facilitate enzymological and biological investigations of this enzyme.
含有六组氨酸亲和前导肽的小鼠C-5胞嘧啶DNA甲基转移酶（MTase，E.C.2.1.1.37）已以比先前报道的水平至少高50倍的水平表达。重组酶的活性水平与野生型酶相似。重组多肽可与镍亲和树脂（IMAC树脂）结合并从其上洗脱。未观察到野生型酶和重组酶在翻译后修饰上有显著差异。该重组系统将有助于进行定点诱变，并将促进对该酶的酶学和生物学研究。
