人突变CD59基因表达蛋白功能的研究

①目的构建8种突变CD59基因真核表达系统，观测突变的人CD59基因在中国仓鼠卵巢（CHO）细胞中的表达及抗补体活性。②方法以荧光活化细胞分类法、免疫荧光法及免疫印迹法检测突变CD59基因转染CHO细胞表达的情况，染料释放实验检测正常突变的CD59分子糖基化前后的抗补体活性。③结果转人突变CD59基因组荧光阳性细胞百分率明显高于对照组，染料释放率低于对照组。④结论CD59突变基因可在CHO细胞表面得到有效表达，该CD59分子具有抗补体活性，但在高糖环境下易被糖基化失活而失去抗补体活性。

A STUDY ON THE FUNCTION OF MUTANT HUMAN CD59 GENE EXPRESSION PROTEIN

Objective To construct eight kinds of mutant CD59 gene expression cell lines and to study the expression and anti complement function of the gene molecules in CHO cell. Methods FACS, FITC and Western-blot were used to confirm transfection and expression of mutant CD59 gene in CHO cells. Dye Release Assay was used to detect the anti-complement function of normal and mutant CD59 molecules before and after glycosylation. Results The fluorescence positive cell rate was significantly higher in the CHO group which had been transfected with mutant CD59 gene than in the control group. The dye release rate was lower than control group. Conclusion Mutant human CD59 genes obtain effectively expression in CHO cell surface. The mutant CD59 molecule bear the anti-complement function, but are easy to lose by glycation in the environment containing high concentration of glucose.