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。
【英文摘要】
To abtain the expression product of subgroup A avian leukosis virus env gene,the env gene amplified from subgroup A avian leukosis virus strain AH10 was cloned into transfer vector p Fast Bac1 and cloned into the shuttle vector Bacmid by transferred into DH10 Bac competent cell. The recombinant shuttle plasmid r Bacmid-env A 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 re...
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