IBDV VP2/VP243基因DNA疫苗表达质粒的构建与表达

Construction and Expression of IBDV VP2/VP243 Genes DNA Vaccine Expressing Plasmids

将传染性法氏囊病病毒 ( IBDV) VP2 /VP2 4 3插入真核表达载体 pc DNA中 ,置于 CMV启动子的调控之下 ,构建成 DNA疫苗表达质粒 pc DNA VP2和 pc DNA VP0。分别转染 CEF细胞后 ,于 2 4、4 8、72 h取细胞裂解液进行 ELISA、SDS-PAGE和 Western blot检测 ,pc DNA VP2于转染后 2 4 h呈阳性反应 ,4 8h表达量高于 2 4 h;pc DNA VP0于转染后 4 8h呈阳性反应 ,72 h表达量高于 4

Two DNA vaccine eukaryotic expressing plasmids,pcDNA VP2 and pcDNA VP0,were constructed by inserting VP2/VP243 genes of IBDV into eukaryotic expressing vector,pcDNA3,which were regulated by the CMV immediate early promoter and enhancer,and stopped by bovine growth hormone (BGH) polyA signal sequence The purpose proteins,VP2 and VP243,could be detected from lysates of transfected CEF cells after 24 h and 48 h of transfection,respectively,with ELISA The results of SDS PAGE and Western blot showed that pcDNA VP2 and pcDNA VP243 could express purpose proteins in CEF cells in vitro ,respectively,which responded specifically to anti IBDV polyclone antibody