[小鼠乳酸脱氢酶C亚基在大肠杆菌中的表达与鉴定]
[Expression and characterization of subunit C of mouse lactate dehydrogenase in Escherichia coli].
作者信息
Xiong Yongzhong, Zheng Dezhu, Xie Fei, Tu Xiangdong, Lan Fenghua
机构信息
Center for Laboratory Medicine, Fuzhou General Hospital of Nanjing Command, PLA, Fuzhou, Fujian 350025, China.
出版信息
Zhonghua Nan Ke Xue. 2004 Jan;10(1):9-11.
OBJECTIVES
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.
目的
构建小鼠乳酸脱氢酶-C的原核重组载体，并检测其在BL21中的表达。
方法
用特异性引物从小鼠睾丸RNA中扩增小鼠乳酸脱氢酶亚基C的编码序列，经BamH I和EcoR I双酶切后克隆到pGEX-2T载体中。用IPTG诱导表达GST融合蛋白。
结果
重组质粒的测序和酶切鉴定表明小鼠乳酸脱氢酶亚基C编码序列存在。IPTG能诱导重组质粒表达出一条约60000的蛋白条带。
结论
小鼠乳酸脱氢酶亚基C的编码序列已导入pGEX-2T质粒，并能高效诱导表达GST融合蛋白。
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