全长及缺失VLDL受体基因转染的CHO细胞与β-VLDL的结合效应

Analysis of the Binding Ability of the Normal VLDL Receptor and its Mutant

为探讨 VLDL受体结合域中 8个重复序列在结合 VLDL中所起的作用 ,利用构建的全长VLDL受体 c DNA和缺失 5个重复序列的该受体 c DNA重组表达载体分别导入 CHO细胞中 .RT- PCR可检测到外源性 VLDL受体基因的表达 .受体与配体结合研究表明 ,转染全长 VLDLR重组体的 CHO细胞结合β- VLDL的能力明显高于转染 VLDLR缺失重组体的 CHO细胞 ,表明人VLDL受体在 CHO细胞中能有效表达 ,而缺失 5个重复序列的

To discuss the contribution of each repeat to ligand binding,two recombinants were constructed,which contained the full length cDNA of VLDLR and the mutant lacking repeats 1-5.The expression of VLDLR gene could be detected by RT PCR from the CHO cells transfected with pCD VR.The results of binding experiments showed that the ability of the CHO cells transfected with the full length cDNA of VLDLR to bind β VLDL was higher than that of the CHO cells transfected with the mutant.The findings indicated that human VLDLR gene could express effectively in CHO cells,and the receptor was almost inactivated when repeats 1-5 were deleted.