摘要
将人源Sef蛋白胞内区编码序列与GST融合构建原核表达质粒进行重组蛋白的表达与纯化并制备多抗 .在COS 7细胞中转染表达hSef显示 ,其分子量分别为 80kD ,10 0kD ,比体外翻译的分子量偏大 ,提示可能有糖基化存在 .Northern印迹的结果表明 ,hSefmRNA主要分布在人肾和睾丸组织 .RT PCR检测到hSefmRNA在众多细胞系有广泛存在 .免疫组化的结果显示 ,hSef蛋白在人肾和睾丸及相应癌组织表达水平较高 .
GST-hSef fusion protein was expressed in E.coli and purified, which was used for the preparation of polyclonal antibody from rabbit. hSef expressed in COS-7 cells showed molecular weight of 80 kD and 100 kD respectively, slightly larger than that from in vitro translation result, which inferred the glycosylation at the potential N-linked glycosylation sites in the extracellular domain. Northern blot showed hSef mRNA was mainly expressed in human kidney and testis tissues. RT-PCR analysis showed a wide spread expression pattern in several cell lines. Immunohistochemical analysis revealed high expression level of hSef protein in kidney, testis and the corresponding carcinoma tissues.The results provide foundation for further study of Sef's function in pathophysiological conditions.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2005年第2期197-203,共7页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家自然科学基金项目 (No.3 0 0 70 70 3
No .3 9970 3 69
No .3 0 0 3 0 0 5 0 )
北京市科学基金项目 (No .H0 2 0 2 2 0 0 2 0 3 10 )联合资助~~
