%0 Journal Article
%T Studies on Establishment of Co-expression System and Function of Human Complement Regulatory Proteins MCP and CD59<br>人补体调节蛋白MCP、CD59共表达体系的构建及其功能研究
%A XU Li
%A ZHAO Zhou-zhou
%A LIU Hui
%A LI Wen-xin
%A <br>徐莉
%A 赵舟宙
%A 刘辉
%A 李文鑫
%J 中国生物工程杂志
%D 2007
%I 
%X Recombinant expression vector pcDNA3-MCPCD59DP containing human membrane complement regulatory proteins(hCRPs) MCP and CD59 cDNA was constructed successfully by using two independent promoters.After transfected into NIH3T3 cells with calcium phosphate-DNA precipitate method,NIH3T3 pcDNA3-MCPCD59-DP transfectants were obtained by G418 selection.Extraneous genes integration was identified by PCR.The co-expression of human CD59 and MCP at both mRNA and protein levels was confirmed by using RT-PCR and Western blot analysis.Human MCP and CD59 cDNA were integrated in NIH3T3 pcDNA3-MCPCD59-DP genomic DNA after continuous 30 times passages,indicating that NIH3T3 pcDNA3-MCPCD59-DP were stable cell lines.Human complement-mediated cytolysis assays showed that NIH3T3 cells transfected stably with pcDNA3-CD59,pcDNA3-MCP,and pcDNA3-MCPCD59-DP were protected from C-mediated damage and co-expressed human MCP and CD59 provided more excellent protection against C-mediated attack as compared with either CD59 or MCP expressed alone.The dicistronic vector represents an effective and efficacy strategy to overcome C-mediated damage and has potential therapeutic value for effectively controlling complement activation and finally for preventing hyperacute rejection in clinical gene therapy.
%K Human complement regulatory proteins Dicistronic vector Co-expression Hyperacute rejection<br>人补体调节蛋白
%K 双基因重组表达载体
%K 共表达
%K 超急性排斥反应
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=951380788B355DEA7D75520FA3B199C9&aid=16B0BFA3AC2BDDD9&yid=A732AF04DDA03BB3&vid=DB817633AA4F79B9&iid=9CF7A0430CBB2DFD&sid=5D311CA918CA9A03&eid=FC0714F8D2EB605D&journal_id=1671-8135&journal_name=中国生物工程杂志&referenced_num=1&reference_num=20