MUC1/Y胞外段在大肠杆菌中的表达和鉴定  

Identification of MUC1/Y extracellular domain expressed in E.coli

作　　者：孙琳[1] 吴秀丽[1] 许艳[1] 于永利[1] 王丽颖[1] 

机构地区：[1]吉林大学基础医学院分子生物学教研室,长春130021

出　　处：《中国免疫学杂志》2006年第9期805-807,811,共4页Chinese Journal of Immunology

摘　　要：目的:获得MUC1/Y胞外段重组蛋白,研究其生物学功能,为肿瘤治疗提供实验依据。方法:利用RT-PCR从MCF7细胞中获得MUC1/Y胞外段编码基因,将其克隆到原核表达载体pET-32a中,并在BL21(DE3)大肠杆菌中进行表达;以亲和层析法对MUC1/Y重组蛋白进行纯化;利用纯化的MUC1/Y蛋白免疫家兔制备MUC1/Y多克隆抗体,然后对乳腺癌组织进行组化染色。结果:在大肠杆菌BL21(DE3)中成功表达了分子量为30000的Trx-MUC1/Y融合蛋白,经镍亲和层析一步纯化所获得的蛋白质纯度>90%,用Trx-MUC1/Y融合蛋白免疫家兔获得的抗血清对乳腺癌组织的初步组化检测证明MUC1/Y融合蛋白具有很好的生物学活性。结论:成功表达并纯化了具有生物学活性的MUC1/Y胞外段重组蛋白。Objective ：To obtain MUC1/Y recombinant protein for further research on its biological function and tumor therapy. Methods：MUC1/Y extracellular domain amplified from MCF7 by RT-PCR was cloned into pET-32a, the MUC1/Y protein was expressed in BL21 （DE3） and then purified by Ni-affinity chromatography;the anti-MUC1/Y polyclonal antibody was gotten from rabbit immunized with MUC1/Y protein and then used to detect breast cancer tissue byimmunohistochemistry. Results：The Trx-MUC1/Y fusion protein（30 000） was successfully expressed in BL21 （ DE3 ） and purified by Ni affinity chromatography ,the purity of Trx-MUC1/Y fusion protein was over 90%. Immunohistochemistry detecting breast cancer with antibody against Trx-MUC1/Y fusion protein proved that Trx-MUC1/Y fusion protein was efficient in biology activity. Conclusion： It is successful to express and purification MUC1/Y extracellular recombinant protein with biology activity.

关 键 词：MUC1/Y胞外段 表达 鉴定 

分 类 号：Q78[生物学—分子生物学]