Expression and characterization of mouse lactate dehydrogenase subunit C in Escherichia coli.

摘要

Objective: To construct a prokaryotic recombinant vector for mouse lactate dehydrogenase-C and to detect its expression in BL21. Methods: The coding sequence of mouse lactate dehydrogenase subunit C was amplified from mouse testis RNA with specific primers and cloned into pGEX-2T after restriction digestion with BamH I and EcoR I. GST fusion protein was expressed after induction with IPTG. Results: Sequencing and restriction digestion of the recombinant plasmid revealed the coding sequence for mouse lactate dehydrogenase subunit C. A protein band of about 60 000 could be induced by IPTG in the recombinant plasmid. Conclusion: 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.