Expression and characterization of subunit C of mouse lactate dehydrogenase in Escherichia coli
Xiong, Y.; Zheng, D.; Xie, F.; Tu, X.; Lan, F.
Zhonghua Nan Ke Xue 10(1): 9-11
2004
ISSN/ISBN: 1009-3591 PMID: 14979197 Document Number: 576532
To construct a prokaryotic recombinant vector for mouse lactate dehydrogenase-C and to detect its expression in BL21. 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. 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. 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.