中国畜牧兽医2012，Vol.39Issue(10)：53-57,5.
猪圆环病毒2型DY株ORF2基因在Sf9昆虫细胞中的表达
Expression of ORF2 Gene of Porcine Circovirus Type 2 in Sf9 Insect Cells
摘要
Abstract
The ORFZ gene of porcine circovirus type 2CPCV2) including complete open reading frame(ORF)was amplified by PCR,and cloned into the pFastBaclTMvector of Bac-to-Bac baculovirus expression systems to construct the recombinant transfer vector pFast-ORF2. The transfer vector was transformed into DHlOBac competent cell which contained the BmNPV bacmid. A recombinant shuttle vector which contained ORF2 gene was constructed by site-specific transposition and the colonies containing recombinant bacmid were collected by white selection. The cultured Sf9 cells were transfected with recombinant bacmid DNA mediating with cellfectin reagent and the pure recombinant baculovirus Bacmid-ORF2 was obtained. After trans-fecting this transformant into Sf9 cells,Cap protein expression in Sf9 cells was detected by indirect immunofluorescent assay and Western blotting. The experimental results showed that the recombinant baculovirus was obtained and the Cap protein with a molecular weight of 31 ku was expressed in Sf9 insect cells. The recombinant protein could be recognized by PCV2 positive serum. This would lay a foundation for developing PCV2 subunit vaccine and diagnosis method.关键词
猪圆环病毒2型/ORF2基因/Sf9昆虫细胞/表达Key words
porcine circovirus type 2/ ORFZ gene/ Sf9 insect cells/ expression分类
生物科学引用本文复制引用
王文秀,沈志强,王善辉,张松林,池贤凤,夏小静..猪圆环病毒2型DY株ORF2基因在Sf9昆虫细胞中的表达[J].中国畜牧兽医,2012,39(10):53-57,5.基金项目
山东省优秀中青年科学家科研奖励基金(BS2011SW026). （BS2011SW026）
