摘要
以杆状病毒Bac-to-Bac表达系统表达猪瘟病毒(CSFV)石门株的E2蛋白,将CSFV石门株的E2囊膜糖蛋白基因亚克隆到杆状病毒转移载体pFastBacHTA中,获得重组转移质粒pFastBacHTA-E2。转化大肠埃希菌DH10Bac感受态细胞,获得重组Bacmid质粒后转染Sf9昆虫细胞,获得重组病毒。传毒3代后对表达蛋白进行Western blot和间接免疫荧光试验检测。结果显示,E2蛋白获得高效表达,能被抗E2蛋白的单克隆抗体2B10和6×His-单克隆抗体特异性识别,表明CSFV石门株E2囊膜糖蛋白在Sf9昆虫细胞中得到成功表达,具有良好的抗原反应性。
Expression of E2 protein of CSFV Shimen strain in baculovirus Bac-to-Bac expression system provided materials for research of the function of E2 protein and develope novel vaccine against CSFV.E2 gene of CSFV Shimen strain was subcloned into a baculovirus transfer vector pFastBacHTA to yield the recombinant transfer plasmid pFastBacHTA-E2.Recombinant bacmid was obtained after transforming the E.coli DH10Bac competent cells.Subsequently,the recombinant bacmid was transfected into Sf9 cells,then got the recombinant virus.The expressed recombinant protein was analyzed by Western blot and indirect immunofluorescence test.The results indicated that E2 protein was efficiently expressed,and had a specific reaction with monoclonal antibody 2B10 and 6×His monoclonal antibody.The E2 protein of CSFV Shimen strain was successfully expressed in Sf9 cells,and had a good antigenicity.
出处
《动物医学进展》
CSCD
北大核心
2012年第1期28-32,共5页
Progress In Veterinary Medicine
基金
浙江省自然科学基金(Y3110396)
