Escherichia coli Expression of Lactate Dehydrogenase C Gene of Black-Lipped Pika in Western Sichuan Plateau
Escherichia coli Expression of Lactate Dehydrogenase C Gene of Black-Lipped Pika in Western Sichuan Plateau作者机构：Department of Lab Medicine Chengdu Medical College Chengdu 610083 China College of Life Science & Technology Southwest University for Nationalities Chengdu 610041 China Animal Husbandry and Veterinary Bureau of Aba County Aba 624000 China
出 版 物：《Animal Husbandry and Feed Science》 (动物与饲料科学（英文版）)
年 卷 期：2011年第3卷第2期
页 面：15-17页
学科分类：0710[理学-生物学] 07[理学] 08[工学] 09[农学] 071007[理学-遗传学] 0901[农学-作物学] 0836[工学-生物工程] 090102[农学-作物遗传育种] 071002[理学-动物学]
基 金：funded by the Applied Fundamental Study of Sichuan Province ( 2008JY0068) China Natural Science Foundation ( 31071700) Funds of the State Ethnic Affairs Commission of PRC ( 08XN04)
主 题：Black-lipped pika Lactate dehydrogenase C Prokaryotic expression
摘 要：[ Objective] To express lactate dehydrogenase C （LDH-C） gene in prokaryotic system and then purify the recombinant protein. E Meth- od] The LDI-I-C gene was amplified from black-lipped pika（ Ochotona curzoniae） by RT-PCR and ligated to the expression vector pET-32a. The E. coli BL21 （DE3） carrying the LDI-I-C gene was obtained and induced by IPTG （isopropy-beta-D-thiogalactoside）. The expression products were analyzed by SDS-PAGE and purified by affinity chromatography. L Result] An approximately 1.0 kbp band appeared in the RT-PCR products as the- oretically expected. As evidenced by PCR and double enzyme digestion, an approximately 1.0 kbp fragment appeared, which indicated successful construction of expression vector. As analyzed by SDS-PAGE, a fusion protein with molecular weight slightly larger than 45 kDa was expressed in form of inclusion body. And very pure fusion protein was obtained by nickel ions-charged affinity chromatography. [ Coaclusion] The LDH-C gene of black-lipped pika has been cloned and expressed in E. coil
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