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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 .
The coding sequence of human Sef protein intracellular region was fused with GST to construct the prokaryotic expression plasmid for the expression and purification of the recombinant protein and the preparation of polyclonal antibody.The expression of hSef in COS 7 cells showed that the molecular weight of the recombinant protein was 80 kD and 100 kD respectively In vitro translation of the molecular weight is large, suggesting the possible presence of glycosylation.Northern blotting results show that, hSefmRNA mainly in human kidney and testicular tissue.RT PCR detected hSefmRNA widespread in many cell lines.Immunohistochemistry results show , hSef protein expression in human kidney and testicular and corresponding high levels of cancer.