Expression and characterization of subunit C of mouse lactate dehydrogenase in Escherichia coli.
- Author:
Yongzhong XIONG
1
;
Dezhu ZHENG
;
Fei XIE
;
Xiangdong TU
;
Fenghua LAN
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Escherichia coli; genetics; Glutathione Transferase; genetics; Isoenzymes; genetics; L-Lactate Dehydrogenase; genetics; Male; Mice; Recombinant Fusion Proteins; biosynthesis; Spermatozoa; enzymology
- From: National Journal of Andrology 2004;10(1):9-11
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVESTo construct a prokaryotic recombinant vector for mouse lactate dehydrogenase-C and to detect its expression in BL21.
METHODSThe 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.
RESULTSSequencing 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.
CONCLUSIONSThe 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.
