小鼠睾丸特异表达新基因(mtLR1)在大肠杆菌中的表达

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。

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

To obtain recombinant mtLRl 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 1PTG 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 mtLRl is purified and obtained by MBP-affinity chromatograph, which may benefit for preparation of specific antibodies against mtLRl protein and its biological activity analysis.