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以ＰＣＲ技术扩增含有ＰｒｅＣ信号肽序列及完整的ＨＢｅＡｇ基因的序列（即ＨＢｃＡｇ基因５′端４４７ｂｐ），在５′端加上合适的酶切位点，克隆到家蚕核多角体病毒转移载体ｐＢｍ０３０上，与野生型ＢｍＮＰＶＤＮＡ共转染家蚕ＢｍＮ细胞，空斑纯化后得到多角体基因失活的重组病毒。ＥＬＩＳＡ法测定表明培养液上清中ＨＢｅＡｇ效价达１∶３２０００，细胞内ＨＢｅＡｇ效价为１∶２０００，培养液及细胞内的ＨＢｃＡｇ含量极低（＜１∶１６０）。研究结果表明，ＢｍＮ细胞能正确识别与切割ＨＢｅＡｇ信号肽序列，所表达的ＨＢｅＡｇ效价高，纯度好，明显优于大肠杆菌表达系统
The sequence of pre-C signal peptide and complete HBeAg gene (ie, 447bp at the 5 ’end of HBcAg gene) was amplified by PCR and cloned into the silkworm nuclear polyhedrosis virus The vector pBm030 was co-transfected with wild-type BmNPVDNA to BmN cells, and the polyhedrin gene-inactivated recombinant virus was obtained after plaque purification. ELISA assay showed that the HBeAg titer in the culture supernatant reached 1:32000, the intracellular HBeAg titer was 1: 2000, and the HBcAg content in the culture solution and cells was very low (<1: 160). The results show that BmN cells can correctly identify and cut the HBeAg signal peptide sequence, the expression of HBeAg titer, good purity, significantly better than the E. coli expression system