摘要
为获得A亚群禽白血病病毒(ALV-A)env基因的表达产物,以ALV-A AH10毒株病毒RNA为模板,通过反转录PCR扩增得到env基因,将其克隆到转移载体p Fast Bac1中,转化DH10Bac感受态细胞,获得重组穿梭质粒r Bacmid-env A,r Bacmid-env A转染昆虫细胞Sf9,获得重组杆状病毒。间接免疫荧光试验(IFA)和免疫印迹试验(Western-blot)结果显示,重组蛋白可与抗ALV-A抗体发生特异性反应,重组蛋白分子大小约为90 000,与预期大小相符,表明env基因在Sf9细胞中获得了良好表达。
To abtain the expression product of subgroup A avian leukosis virus env gene,the env gene amplified fromsubgroup A avian leukosis virus strain AH10 was cloned into transfer vector pFastBaci and cloned into the shuttle vector Bacmid by transferred into DH10Bac competent cell. The recombinant shuttle plasmid rBacmid-envA was then transfected into Sf9 cells to generate recombinant virus. The results of indirect immunofluorescence assay （ IFA） and western blot showed that the recombinant protein could be recognized specifically by the polyclonal antibody against ALV-A with the molecule weight about 90 000,which was consistent with the expected,and all the results indicated that the env gene was expressed well in Sf9 cells .
出处
《江苏农业学报》
CSCD
北大核心
2017年第6期1316-1320,共5页
Jiangsu Journal of Agricultural Sciences
基金
国家自然科学基金项目(31201881)
关键词
A亚群禽白血病病毒
ENV基因
表达
avian leukosis virus subgroup A（ALV-A）
env gene
expression
