(肿瘤生物学)人单酰基甘油脂肪酶基因的克隆表达_纯化及活性研究_朱京童.pdf

DOI:10.13461/ki.cja.005031 . 593 . 1001-8689(2012)08-0593-06 1 2 1 1,* 1 1,* 1 (1 050015 2 610064) (human monoacylglycerol lipase, MAGL) MAGLMAGL RT-PCRRNAMAGLpET28a(+) E. coli BL21(DE3)1mmol/L IPTGNi-NTA SDShydroxycoumarinyl arachidonate (7-HAC) MAGLpET28a-MAGLSDSMAGL 25%Ni-NTA90%7900IU/mg, 23MAGLN-arachidonyl maleimide(NAM)IC500.53μmol/L MAGL R979.1 A Cloning, expression and purifi cation of human monoacylglycerol lipase 1 2 1 1 1 1 1 Zhu Jing-tong , Zhang Shen , Zheng Hai-zhou , Zheng Zhi-hui , Lu Xin-hua , Dong Ai-hua and Duan Bao-ling (1 New Drugs Research Development company of North China Pharmaceutical Group Corporation, National Engineering Research Center of Microbial Medicine, Shijiazhuang 050015; 2 College of Life Science, Sichuan University, Chengdu 610064) Abstract Objective Human monoacylglycerol lipase (MAGL), a serine hydrolase that converts monoglycerides to fatty acid and glycerol, plays crucial roles in cancer pathogenesis, and has become a novel drug target for antitumor drug development. In present study, to develop a high throughput screening asaay for MAGL inhibitor, the human MAGL was recombinantly expressed in E. coli . Methods The human MAGL gene was cloned by reverse transcription-PCR(RT-PCR) from human brain total RNA, and was inserted into expression vector pET28a(+). The recombinant MAGL protein was expressed in E. coli host strain B