Snake Muscle Creatine Kinase cDNA： Molecular Cloning, Expression and Comparison with Evolutionarily Related Enzymes

ZHANG Feng-Wen, YANG Liang^*, ZHAO Fu-Kun, Xu Gen-Jun^**
( Shanghai Institute of Biochemistry, the Chinese Academy of Sciences, Shanghai 200031, China )

Abstract    Snake muscle cDNA library was constructed, and an allelic cDNA coding for M-type creatine kinase was cloned through screening library with antibody. A complete open reading frame(1 140 bp) codes for 380 amino acids which shows high homology with known creatine kinase isoenzyme. After cloning into expression vector pET11a, the snake muscle creatine kinase(SM-CK) was over-expressed in Escherchia coli. Kinetical studies showed that purified recombinant SM-CK exhibited behavior similar to the tissue form. Evolutionary tree of M-type creatine kinase was then constructed, and it was determined that this allelic cDNA of snake muscle creatine kinase belonged to M-type situated between in mammals and birds. These findings, along with the following SM-CK gene regulation studies, may shed light on the role of creatine kinase in energy generation during hibernation as well as the molecular basis of myogenesis during ontogeny.
Key words    Snake muscle creatine kinase； gene cloning and expression； homology； evolutionary tree

^*Graduate student from East China Normal University
^**Corresponding author： Tel, 86-21-64374430-257； Fax, 86-21-64338357； e-mail： [email protected]