小鼠睾丸特异表达新基因(mtLR1)在大肠杆菌中的表达  

Expression and purification of mouse testis-specific gene mtlr1 protein in E. coli

作　　者：聂东宋[1] 周延凯[1] 刘宇[1] 曹佐英[1] 

机构地区：[1]湖南理工学院化学化工系,湖南岳阳414000

出　　处：《湖南理工学院学报（自然科学版）》2006年第4期67-69,89,共4页Journal of Hunan Institute of Science and Technology(Natural Sciences)

基　　金：国家重点基础研究发展规划973项目(G1999055901)

摘　　要：根据已经克隆的mtLR1基因序列,采用RT-PCR的方法获得mtLR1基因。将所得的PCR产物插入原核表达载体pMAL-P-2x中,得重组质粒(pMAL-P-2x/mtLR1)并转化大肠杆菌(E．coli)DH5α．经IPTG诱导表达,SDS-PAGE电泳分析显示,重组蛋白得到了正确表达,表达的融合蛋白占菌体蛋白总量的52％,分子质量约为73 kDa。mtLR1蛋白的高效表达,为研究其生物学功能和制备单克隆抗体奠定了基础。To obtain recombinant mtLR1 protein by prokaryotic expression for further study on its functions, the encoding sequence for mature protein of mouse mtLRl gene was amplified with RT-PCR and inserted into pMAL-p-2x vector to establish the prokaryotic expressing system. The competent cells of host strain of DH5a were transformed by the recombinant plasmid （pMAL-p-2x/mtLR1）. Expression of the target protein was induced with IPTG and assayed by SDS-PAGE after DNA sequencing. The Expressed fusion protein is 73 kD in SDS-PAGE as expected and accounts for 52% of the total bacteria proteins. The recombinant mtLR1 is purified and obtained by MBP-affinity chromatograph, which may benefit for preparation of specific antibodies against mtLRl protein and its biological activity analysis.

关 键 词：mtLR1 表达与纯化 大肠杆菌 

分 类 号：Q492.4[生物学—生理学]