Gq蛋白竟争性抑制肽GCIP-27质粒载体的构建及表达  

Construction and expression of G protein competitive inhibitory polypeptide GCIP-27

作　　者：周见至[1] 李晓辉[1] 吴雪晖[2] 张海港[1] 

机构地区：[1]第三军医大学药学院药剂教研室 [2]第三军医大学西南医院矫形外科,重庆400038

出　　处：《重庆医学》2007年第21期2172-2173,共2页Chongqing medicine

基　　金：重庆市科技攻关重点项目(20048256)

摘　　要：目的化学合成寡核苷酸片段克隆入pIVEX2.3质粒中,构建GCIP-27的原核表达载体并进行原核表达。方法化学合成GCIP-27的全基因序列,定向克隆到pIVEX2.3质粒中构建含有T7启动子的pIVEX2.3-GCIP27原核表达载体并酶切及测序鉴定;pIVEX2.3-GCIP27质粒转染E coli BL21(DE3)pLysE细菌,用IPTG诱导GCIP-27多肽的表达。结果成功构建了pIVEX2.3-GCIP27表达质粒,经测序鉴定结果与设计的完全相符;成功用BL21细菌表达了GCIP-27多肽,GCIP-27占细菌总蛋白的4.96%。结论本实验构建并表达了GCIP-27的重组质粒,为GCIP-27的大量原核表达奠定了基础。Objective To construct and expression a recombinant plasmid for expressing peptides of GCIP 27. Methods Nucleotide fragments were synthetized, and double-stranded oligonucleotide was subcolned into pIVEX2.3 plasmid vector, which was digested with Nco I and Sma I, for expression in E coli BL21（DE3）pLysE. pIVEX2.3-GCIP27 plasmid was identified by DNA sequencing. Recombinant plasmid was expressed induced by IPTG. The expression of GCIP-27 was identified by SDS-PAGE. Results The pIVEX2.3-GCIP27 plasmid was successfully constructed and confirmed by DNA sequencing. The GCIP-27 was expressed in E coli BL21（DE3）, and was about 4.96% . Conclusion With DNA recombination techniques, the expression plasmid, which include the GCIP-27 gene and under the control of T7 promoter,was cloned and expression successsfully.

关 键 词：GCIP-27 心肌肥大 pIVEX2.3 表达 

分 类 号：R541[医药卫生—心血管疾病]