小鼠丝氨酸消旋酶的克隆、表达与纯化  

Gene Cloning,Expression,and purification of the serine racemase

作　　者：段英俊[1] 刘俊林[1] 石亚伟[1] 

机构地区：[1]山西大学生物技术研究所教育部化学生物学与分子工程重点实验室,山西太原030006

出　　处：《生物技术世界》2013年第3期4-5,共2页Biotech World

基　　金：国家自然基金(30400065;31170748)资助

摘　　要：丝氨酸消旋酶(Serine Racemase,SR)是一种磷酸吡多醛依赖酶,在ATP和Mg2+的辅助下,催化L型丝氨酸转变为D型丝氨酸,D-丝氨酸通过结合在NMDA受体的Gly结合位点,调节其生理功能。本文将小鼠来源的丝氨酸消旋酶基因克隆至原核表达载体pMAL-C2上,构建重组质粒pMAL-C2-SR。将重组质粒转入E.coil BL21(DE3)宿主菌中,经IPTG诱导,获得重组表达。重组蛋白带有MBP标签,经Amylose亲合柱和Sephacryl S-200纯化,获得电泳纯的目的蛋白。Serine racemase (SR) is a pyridoxal phosphate dependent enzyme that catalyzes the interconversion of L and D-serine isomers in the presence of ATP and Mg2+ .D-serine play a critical role in regulation of NMDA receptor function by its Gly binding site.Serine racemase gene from mice was cloned into pMAL-C2 to construct recombinant plasmid pMAL-C2-SR.The recombinant plasmid pMAL-C2-SR was transformed into E.coli BL21 and expressed by isopropyl-β-d-thiogalactoside (IPTG) induction.The fusion protein with MBP tag was purified by Amylose column and Sephacryl S-200 column.

关 键 词：丝氨酸消旋酶 D-丝氨酸 克隆 纯化 

分 类 号：Q78[生物学—分子生物学]