乳腺珠蛋白基因在大肠杆菌中的诱导表达及纯化研究  

Prokaryotic Expression and Purification of Human Mammaglobin Gene in E.coli

作　　者：刘秀娜[1] 王仙园[1] 周娟[1] 潘自铁[2] 赵力军 胡川闽[2] 

机构地区：[1]第三军医大学护理管理学教研室 [2]第三军医大学临床生化教研室,重庆400038 [3]National Centerfor HIV,STD and TB Prevention,Centers for Disease Control and Prevention

出　　处：《解放军护理杂志》2005年第11期7-9,44,共4页Nursing Journal of Chinese People's Liberation Army

基　　金：重庆市科委应用基础研究资助项目(2004);校归国人员启动资金资助(2004)

摘　　要：目的克隆乳腺珠蛋白(hum an m amm aglob in,hMAM)编码全长cDNA,原核表达并纯化蛋白产物,为后期研制乳腺癌早期诊断试剂盒奠定基础,从而为早期发现乳腺癌提供科学的监测方法。方法自乳腺癌组织提取总RNA,通过RT-PCR克隆hMAM cDNA,构建pQE40-hMAM表达质粒,在大肠杆菌M15中表达,利用镍-亚硝胺乙酸组氨酸(N i-NTA-H is)亲和层析法对重组蛋白进行纯化。结果表达的融合蛋白以不溶性包涵体形式存在,纯化后得到纯度为97%的目的蛋白。结论成功地纯化出hMAM重组蛋白。Objective To clone the cDNA in full-length of human mammaglobin （hMAM） and to do prokaryotic expression and purify hMAM protein, in order to provide an experimental basis for early diagnostic reagent bottle and early screening of breast cancer. Methods hMAM cDNA was amplified through RT-PCR from breast cancer tissue. The recombination pQE40-hMAM vector was constructed and expressed in E. coli M15 after induction by IPTG. The fusion protein was purified with Ni-NTA-His affinity chromatography. Results The fusion protein formed inclusion body in prokaryotic expression system and the renatured protein was purified with purity of 97%. Conclusion The recombinant hMAM is successfully purified.

关 键 词：乳腺癌 人乳腺珠蛋白 原核表达系统 蛋白纯化 

分 类 号：R730.54[医药卫生—肿瘤] R737.9[医药卫生—临床医学]