人IL-16在大肠杆菌中的融和表达  

Fusion expression of human interleukin-16 in E. coli

作　　者：曹廷兵[1] 叶治家[1] 甘立霞[1] 钟小林[1] 巩燕[1] 彭家和[1] 

机构地区：[1]第三军医大学基础医学部生物化学与分子生物学教研室,重庆400038

出　　处：《第三军医大学学报》2002年第4期428-430,共3页Journal of Third Military Medical University

基　　金：全军"九五"医药卫生基金资助项目 ( 96L0 31);重庆市攻关项目( 970 4 3)

摘　　要：目的　将人IL 16cDNA克隆至原核融合表达载体PinpointTMXa 3上 ,并进行表达研究。方法　含IL 16cDNA的质粒IL 16/PGEM 3ZF(+ )和PinpointTMXa 3原核融合表达载体经酶切消化、目的DNA片段的分离、重组连接、筛选、序列分析等系列操作 ,获得重组质粒PinpointTMXa IL 16,将质粒PinpointTMXa IL 16转化至大肠杆菌JM10 9,用IPTG诱导表达JM10 9工程菌 ,Westernblotting检测表达产物。结果　IL 16cDNA成功克隆至融合表达载体PinpointTMXa 3 ,Westernblotting检测经诱导含有PinpointTMXa IL 16质粒的JM10 9细菌 ,有IL 16融合蛋白的表达 .结论　成功地表达了IL 16融合蛋白。Objective To construct the plasmid Pinpoint TM Xa IL 16 and then observe the expression of fused IL 16 in E.coli . Methods IL 16 cDNA was isolated from plasmid IL 16/PGEM 3Zf(+) and ligated to the expression vector Pinpoint TM Xa 3 after enzyme digestion, target DNA fragment isolation, recombinant ligation, and screening. After the DNA sequence of plasmid Pinpoint TM Xa IL 16 was analyzed, it was transfected to E. coli strain JM109 with the inducement of IPTG. The expression of IL 16 was identified with Western blotting. Results Pinpoint TM Xa IL 16 was constructed successfully and IL 16 fused with the biotin was expressed in E. coli . Conclusion IL 16 is expressed in E. coli , which found a basis for batch production of the fusion protein.

关 键 词：IL-16 克隆 融合表达 大肠杆菌 

分 类 号：R392.12[医药卫生—免疫学] R378.21[医药卫生—基础医学]