脊髓损伤修复相关10号蛋白在COS7细胞中的表达、纯化与鉴定  

Expression of Spinal Cord Injury and Regeneration Related Protein 10 in COS7 Cells and its Purification and Identification

作　　者：唐宁[1] 阙海平[1] 刘少君[1] 

机构地区：[1]军事医学科学院基础医学研究所,北京100850

出　　处：《生物技术通讯》2010年第5期637-640,共4页Letters in Biotechnology

基　　金：国家高技术研究发展计划(2007AA02Z183)

摘　　要：目的:在COS7细胞中表达脊髓损伤修复相关10号蛋白(SCIRR10),并对表达产物进行纯化和鉴定。方法:在实验室前期研究基础上,用PCR方法扩增SCIRR10基因,将其克隆入pcDNA3.1/myc-HisA穿梭质粒中,转染COS7细胞进行表达;对表达产物用金属螯合层析方法纯化,并进行SDS-PAGE和Western印迹检测。结果及结论:构建了含有SCIRR10基因的穿梭质粒pcDNA3.1/myc-His-SCIRR10,并使其在COS7细胞中得到表达,纯化后的重组蛋白SCIRR10-His-myc的纯度在80%以上,可用于下一步的实验研究。Objective： To carry out the expression of spinal cord injury and regeneration related protein No.10（SCIRR10） in COS7 cells,and to purify and identify the recombinant protein.Methods： The gene of SCIRR10 which was got by researchers in our laboratory,was cloned into E.coli-eukaryotic cell shuttle plasmid pcDNA3.1 / myc-HisA and transfected into COS7 cells.The expressing products were purified by metal chelate chromatography.Then,the purified protein was analyzed by SDS-PAGE and Western blot.Results Conclusion： A recombinant shuttle plasmid pcDNA3.1 / myc-His-SCIRR10 was successfully constructed and SCIRR10 was successfully expressed in COS7 cells.SDS-PAGE and Western blot analysis showed that purified recombinant protein had purity of more than 80%.This work provides the basis for further study on the function of SCIRR10.

关 键 词：脊髓损伤修复相关10号蛋白 真核表达 COS7细胞 金属螯合层析 

分 类 号：Q78[生物学—分子生物学]