Protein trans-splicing based dual-vector delivery of the coagulation factor Ⅷ gene

查看全文 作　　者：ZHU FuXiang,LIU ZeLong,CHI XiaoYan & QU HuiGe Life Science College of Ludong University,Yantai 264025,China

出　　处：《Science China(Life Sciences)》索引2010年第53卷第6期,共7页

基　　金：supported by the Natural Science Foundation of Shandong Province, China (Grant No. Y2005D14);the Science and Technology Program of Yantai (Grant No. 2008152);the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry (Grant No. 20071108);the Discipline Construction Funds of Ludong University 摘　　要：A dual-vector system was explored for the delivery of the coagulation factor VIII gene,using intein-mediated protein trans-splicing as a means to produce intact functional factor VIII post-translationally.A pair of eukaryotic expression vectors,expressing Ssp DnaB intein-fused heavy and light chain genes of B-domain deleted factor VIII (BDD-FVIII),was constructed.With transient co-transfection of the two vectors into 293 and COS-7 cells,the culture supernatants contained (137±23) and (109±22) ng mL–1 spliced BDD-FVIII antigen with an activity of (1.05±0.16) and (0.79±0.23) IU mL–1 for 293 and COS-7 cells,respectively.The spliced BDD-FVIII was also detected in supernatants from a mixture of cells transfected with inteinfused heavy and light chain genes.The spliced BDD-FVIII protein bands from cell lysates were visualized by Western blotting.The data demonstrated that intein could be used to transfer the split factor VIII gene and provided valuable information on factor VIII gene delivery by dual-adeno-associated virus in hemophilia A gene therapy. 关键词：INTEIN protein TRANS-SPLICING coagulation factor VIII dual-vector GENE delivery

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Protein trans-splicing based dual-vector delivery of the coagulation factor Ⅷ gene

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