The Study On The Relationship Between Mutant CD59 And Vescular Proliferation Of Diabetes

Objective TO amplify two human mutant CD59 eukaryotic expressing system and investigate mutant CD59 functional activity.Methods Mammalian expression vector PLATER of mutant CD59 cDNAs was transfected into CHO together with the pcDNA by lipofectamine,which confers resistance to G418. The positive clones were tested by FIH and EIH and FACS. Activity of both mutants CD59 was determined by BCECF release assay.Results A eukaryotic system that expressing human mutant CD59 cDNA was successfully set up.Mutant CD59 cDNAs subcloned into the mammalian expression vector PLATER and transfected into CHO together with the pcDNA,with confers resistance to G418. The positive clones were tested by FIH and EIH.FACS indicated these expression rates of two mutant CD59 on CHO cells were 36.6%,41.7%.Activity of both mutants CD59 before and after glycation was determined by BCECF release assay,both of them could restrict MAC formation ,and glycation could inhibit CD59.Conclusion A eukaryotic system that expressing mutant CD59 cDNA was successfully set up.We investigated mutant CD59 could restrict MAC formation,and glycation could inhibit mutant CD59. These would be helpful for the furthur study of link mutant CD59 and the vascular proliferative of diabetes.