麻疯树肌醇-1-磷酸合成酶基因的克隆与原核表达  

Molecular cloning and prokaryotic expression of a novel 1L-myo-inositol-1-phosphate synthase gene from Jatropha curcas

作　　者：喻川[1] 苟春宝[1] 黄静[1] 王勇[1] 陈放[1] 魏炜[1] 

机构地区：[1]四川大学生命科学学院生物资源与生态环境教育部重点实验室,四川省生物资源与生物制药重点实验室,成都610065

出　　处：《四川大学学报（自然科学版）》2011年第6期1397-1402,共6页Journal of Sichuan University(Natural Science Edition)

基　　金：“十一五”科技支撑项目(2006BAD07A04);四川省“十一五”支撑项目(2007BAD50800)

摘　　要：通过RACE技术,克隆了麻疯树肌醇-1-磷酸合成酶基因(JcMIPS),其开放读码框为1530 bp,编码510个氨基酸.序列分析表明,JcMJPS与多种植物MJPS基因的氨基酸序列具有较高相似性,达87.08%～91.18%,且含有MIPS基因的四个保守域.在此基础上构建了原核表达栽体,在1 mmol/L IPTG,18℃下诱导表达5 h,获得了可溶性的目的蛋白.亲和层析纯化目的蛋白,通过体外酶学反应鉴定,纯化样品具有酶学活性,表明JcMIPS原核表达成功.The 1L-myo-inositol-l-phosphate synthase gene of Jatropha curcas was cloned by RACE tech- nique. Its open reading frame is 1530 bp, encoding 510 amino acids. Amino acid sequences analysis showed that JcMIPS had high similarity with MIPS genes of multiple plants, up 87.08M to 91.18M, and contained four conserved domains of the MIPS gene. The prokaryotic expression vector was constructed and was induced to express in 1 mmol/L IPTG and 18℃ for 5 hours to obtain a soluble protein. The purified target protein was identified by enzymatic reactions in vitro, suggesting that the sample had the enzyme activity and indicating the success of JcMIPS prokaryotic expression.

关 键 词：麻疯树 肌醇-1-磷酸合成酶 克隆 原核表达 

分 类 号：Q94[生物学—植物学]