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


CONCLUSIONS
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.
Center for Laboratory Medicine, Fuzhou General Hospital of Nanjing Command, PLA, Fuzhou, Fujian 350025, China.
Yongzhong
Y
Xiong
Dezhu
D
Zheng
Fei
F
Xie
Xiangdong
X
Tu
Fenghua
F
Lan
D000818:Animals; D004926:Escherichia coli / Q000235:genetics*; D005982:Glutathione Transferase / Q000235:genetics; D007527:Isoenzymes / Q000235:genetics*; D007770:L-Lactate Dehydrogenase / Q000235:genetics*; D008297:Male; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D013094:Spermatozoa / Q000201:enzymology*
D007527:Isoenzymes; D011993:Recombinant Fusion Proteins; D007770:L-Lactate Dehydrogenase; C409673:lactate dehydrogenase C4; D005982:Glutathione Transferase