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目的应用新型杆状病毒表达系统快速构建含有ＨＢｓＡｇ基因的重组杆状病毒，高效表达ＨＢｓＡｇ，为ＨＢＶ诊断试剂、疫苗及治疗研究提供依据。方法构建含有ＨＢｓＡｇ基因的供体质粒ｐＦＢ－ＢＳ，转化Ｂａｃ－ｔｏ－Ｂａｃ杆状病毒表达试剂盒中的ＤＨ１０Ｂａｃ致敏菌，利用其含有的细菌Ｔｎ７转座系统将ＨＢｓＡｇ基因重组至穿梭质粒Ｂａｃｍｉｄ上，快速筛选出含有ＨＢｓＡｇ基因的重组杆状病毒。结果此重组病毒能在昆虫细胞中表达出有生物学活性的ＨＢｓＡｇ，表达产物主要集中在细胞内，ＥＬＩＳＡ滴度为１：２×１０４，产量可以达到２μｇ／１０６细胞。结论此系统的成功应用为真核表达工作提供新的工具
OBJECTIVE: To rapidly construct a recombinant baculovirus containing HBsAg gene by using a novel baculovirus expression system to express HBsAg efficiently and provide a basis for HBV diagnostic reagents, vaccines and therapeutic research. Methods The donor plasmid pFB-BS containing HBsAg gene was constructed and transformed into DH10Bac-expressing bacteria in Bac-to-Bac baculovirus expression kit. HBsAg gene was recombined into shuttle plasmid Bacmid using the bacterial Tn7 transposing system , Quickly screened recombinant baculovirus containing HBsAg gene. Results The recombinant virus could express HBsAg in insect cells. The expression products mainly concentrated in the cells. The ELISA titer was 1: 2 × 104 and the yield could reach 2μg / 106 cells. Conclusion The successful application of this system provides a new tool for eukaryotic expression