人胸腺素α_1基因工程菌的构建与表达  

Construction and expression of recombinant human thymosin alpha 1

作　　者：林陈水[1] 黎小军[1] 王平[1] 梅建凤[1] 

机构地区：[1]浙江工业大学药学院,浙江杭州310032

出　　处：《浙江工业大学学报》2005年第3期306-309,共4页Journal of Zhejiang University of Technology

基　　金：浙江省科技厅资助项目(2005C33031)

摘　　要：胸腺素α1(Thymosinalpha1,Tα1)是一种针对T淋巴细胞的免疫增强剂,临床用途广泛.为大量制备Tα1,按大肠杆菌惯用密码子合成编码Tα1的基因,克隆于pUCmT,转化E.coliJF1125,经测序证明序列正确后,克隆入pET39(b)的BspT 和BamH 位点,成功地构建出包含信号肽,DsbA,连接肽,6个His,EKsite(Asp-Asp-Asp-Asp-Lys),胸腺素α1的表达载体,转化E.coliBL21(DE3),获得胸腺素α1基因工程菌.SDS-PAGE分析表明,经0.1mmol/LIPTG和30℃诱导8h,在大肠杆菌周质中大量表达表观分子量31kD左右的融合蛋白.为进一步获得大量可供实验研究和临床应用的重组胸腺素α1创造条件.<Abstrcat> Thymosin alpha 1(Tα_1) is a immunopotentiating for T cell agent widely used in clinic. In order to prepare Tα_1 in large scale, the gene of Tα_1 was synthesized according to the preferential codons of E.coli, then cloned into pUCmT vector and transformed into E.coli JF1125. The target gene was inserted into BspTⅠand BamHⅠ sites of expression vector pET39(b) after sequencing, and the recombinant plasmid was transformed into E.coli BL21(DE3). Successfully constructed the expression plasmid that containing signal peptide, DsbA,linker peptide, 6 His, EKsite(Asp-Asp-Asp-Asp-Lys) and Tα_1.SDS-PAGE analysis showed that higher levels of expression of fusion protein in periplasmic space, with a molecular weight of about 31 kD , induced at 30 ℃ by 0.1 mmol/ L IPTG for 8 hours. It can provide foundation for obtaining a larger quantity of recombinant Tα_1 for experimental and clinic studies.

关 键 词：胸腺素α1(Tα1) 基因克隆 融合表达 

分 类 号：Q789[生物学—分子生物学]