Expression and characterization of subunit C of mouse lactate dehydrogenase in Escherichia coli / 中华男科学杂志

ABSTRACT

<p><b>OBJECTIVES</b>To construct a prokaryotic recombinant vector for mouse lactate dehydrogenase-C and to detect its expression in BL21.</p><p><b>METHODS</b>The coding sequence of mouse lactate dehydrogenase subunit C was amplified from mouse testis RNA with specific primers, and cloned into pGEX-2T after the restriction digestion with BamH I and EcoR I. GST fusion protein was expressed after induction with IPTG.</p><p><b>RESULTS</b>Sequencing and restriction digestion of the recombinant plasmid revealed the existence of coding sequence for mouse lactate dehydrogenase subunit C. A protein band of about 60,000 could be induced by IPTG in the recombinant plasmid.</p><p><b>CONCLUSIONS</b>The coding sequence of mouse lactate dehydrogenase subunit C was introduced into the pGEX-2T plasmid and a GST-fused protein could be induced at a high level.</p>