鹅细小病毒NS2基因以及VP1与VP3非重叠序列基因的真核表达

采用pBlueBacHis2A系统筛选鹅细小病毒(GPV)的检测抗原,利用PCR扩增和双酶切方法分别构建了含有NS2基因和VP1与VP3非重复序列基因的重组质粒pBlueBacHis2A-GPV-NS2、pBlueBa-cHis2A-GPV-VP(1-3),分别转染sf9细胞,得到重组病毒,分别表达出了54 ku的NS2融合蛋白和24 ku的VP(1-3)融合蛋白。Western-blotting和Dot-ELISA检测结果证实,表达产物具有特异性;间接免疫荧光试验证实,重组蛋白在细胞中获得了表达。

Eukaryotic expression of NS2 gene and non-overlapping sequence of VP1 gene to VP3 gene of goose parvovirus and detection of the expressed proteins

The pBlueBacHis2A insect expression system was chosen to screen the effectual detecting antigens against goose parvovirus(GPV).Recombinant plasmids pBlueBacHis2A-GPV-NS2 with NS2 gene of GPV and pBlueBacHis2A-GPV-VP(1-3) with the nonoverlapping sequence of VP1 gene to VP3 gene of GPV were constructed by PCR and double restriction enzymes.Then,the two plasmids were transfected into sf9 cells to obtain recombinant viruses.The expressed proteins were 54 ku and 24 ku in size from the two recombinant plasmids,respectively,and their specificities were detected by Western-blotting,Dot-ELISA and indirect immunofluence assay.All the results confirmed that the recombinant proteins which were expressed successfully in sf9 cells had good specificity.