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目的 采用新型杆状病毒表达系统(Bac to Bac系统)在昆虫细胞Sf-9中进行新城疫病毒F48 E8株HN基因表达。方法NDV F48 E8株 HN基因插入到转座载体 pFast Bac Ⅰ,构建重组转座载体 pFast HN,转化到感受态细胞 DH10后,将目的基因转座到穿梭载体Bacmid中,经筛选获得重组Bacmid HN转染到Sf-9昆虫细胞,并进行检测。结果 表达产物经SDS-PAGE、Westem blot检测获得蛋白特异带,相对分子质量约63000。结论NDVF48E8株HN基因能通过新型杆状病毒表达系统在昆虫细胞Sf-9中进行表达,表达量约为细胞总蛋白的10％。
Objective To express HN gene of Newcastle disease virus F48 strain E8 in insect cell Sf-9 using a novel baculovirus expression system (Bac to Bac system). Methods The HN gene of NDV F48 E8 strain was inserted into the transposase vector pFast Bac Ⅰ to construct the recombinant transposase pFast HN. After transformed into competent cells DH10, the target gene was transplanted into the shuttle vector Bacmid. The recombinant Bacmid HN Sf-9 insect cells were stained and tested. Results The expressed protein was detected by SDS-PAGE and Western blot. The specific molecular weight was about 63000. Conclusion The HN gene of NDVF48E8 strain can express in insect cell Sf-9 through a novel baculovirus expression system, and the expression level is about 10% of the total cellular protein.