Title: EXPRESSION OF HUMAN MUTANT CD59 IN CHINESE HAMSTER OVARY CELLS IN PATIENTS WITH DIABETES
Abstract: Objective To construct a eukaryotic expression bearer of two human mutant CD59 and obtain Chinese hamster ovary(CHO) cell eukaryotic expression system,so as to interpret the mechanism of vascular proliferation in diabetes. Methods pALTER plasmid containing two mutant CD59 cDNAs was transfected into CHO cells together with pcDNA by positive liposome.The positive clones were obtained by G418.The expression of CD59 on the cell membrane was tested by cell immunohistochemistry and flow cytometry. Results pALTER plasmid containing CD59 was cut with restriction enzymes and a 496 bp fragment obtained by electrophoresis,which was complete conformity with the insert.The pALTER plasmid was successfully transfected into CHO cells and positive clones were screened and detected immunohistologically.It was proved that CD59 was able to express on the surface of CHO cell,with the expression rates being 68.6% and 71.7%,respectively. Conclusion Two kinds of high-performance expressions of eukaryotic system of human mutant CD59 cDNA were successfully established,which makes further study of mutant CD59 glycation and anti-complement activity possible,and thus elucidate the mechanism of blood vessel proliferation in diabetes.
Publication Year: 2009
Publication Date: 2009-01-01
Language: en
Type: article
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