J亚群禽白血病病毒env基因主要功能区和EGFP基因在SSff9细胞中的融合表达  

Fusion Expression of Main Function Domain of env Gene of Avian Leukosis Virus Subgroup J and EGFP Gene in Sf9 Cells

作　　者：李拓凡 多婷[1] 梁雄燕[1] 顾玉芳[1] 杨玉莹[1] 

机构地区：[1]长江大学动物科学学院,湖北荆州434025

出　　处：《中国家禽》2015年第11期23-26,共4页China Poultry

基　　金：国家自然科学基金项目(31060342)

摘　　要：为获得具备良好抗原性和生物活性的J亚群禽白血病病毒(ALV-J)env基因主要功能区的表达产物,采用特异性引物分别从保存的p MD18T-envHB2010质粒、PIRES2-EGFP中扩增ALV-J env基因主要功能区和增强型绿色荧光蛋白(EGFP)基因,酶切、连接后插入p Fast Bac Daul载体,构建杆状病毒转移载体p Fast Bac Daul-env-EGFP,将其转化DH10Bac感受态细胞制备重组杆粒Bacmid-env-EGFP,转染Sf9细胞进行真核表达。结果显示,转染了重组杆粒的Sf9细胞在倒置荧光显微镜下呈现亮绿荧光;以ALV-J单克隆抗体JE9进行Western blot检测,转染重组杆粒的Sf9细胞检测出约85 ku的条带。结果表明,目的基因在Sf9细胞中得到了良好的表达,为进一步研究ALV-J提供基础试验材料。The objective of this study was to get the expression product of main function domain of avian leukosis virus subgroup J （ALV-J）with good antigenicity and biological activity. The env gene and enhanced green fluorescent protein（EGFP） gene were cloned into pFastBac Daul vector to construct pFastBac Daul-env-EGFP transfer vector. The recombinant bacmid Bacmid-env-EGFP was constructed through transfering the pFastBac Daul-env-EGFP into DH10Bac competent cell. Sf9 cells which were transfected with Bacmid-env-EGFP showed brightly green fluorescence after 5 days. Positive band was detected through Western blot in the lysate of cells which were transfected. The results suggested that recombined bacmid was expressed in Sf9 cells,which supplied the material for further study of ALV-J.

关 键 词：ALV-J ENV基因 EGFP 融合表达 

分 类 号：S855.3[农业科学—临床兽医学]