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通过ＰＣＲ获得长度为６４０ｂｐ的ＨＢＶｐｒｅＣ／Ｃ基因片段，将其克隆入转移载体质粒ｐＳＸＩＶＶＩ＋Ｘ３／４，构建出重组质粒ｐＳＸＩＶＶＩ＋Ｘ３／４－ｐＣ／Ｃ。利用共转染的方法，构建出既能形成多角体又能表达ｐｒｅＣ／Ｃ基因的重组毒株ＴｎＮＰＶ－ｐＣ／Ｃ－ＯＣＣ＋。该重组毒株中ｐｒｅＣ／Ｃ基因受到串联的双启动子－合成启动子和含ＨｉｎｄⅢ接头的ＸＩＶ启动子的双重调控。对感染重组毒株的草地夜蛾（Ｓｐｏｄｏｐｔｅｒａｆｒｕｇｉｐｅｒｄａ，Ｓｆ）细胞及其培养上清分别进行ＨＢｅＡｇ、ＨＢｃＡｇ固相放射免疫检测，发现受感染细胞及其培养上清均呈ＨＢｅＡｇ阳性，ＨＢｃＡｇ阴性；进一步做Ｗｅｓｔｅｒｎ分析表明，受感染细胞总蛋白中２６ｋＤ及培养上清１５ｋＤ处，呈现与ＨＢｅＡｇ单克隆抗体特异性反应条带。结果表明，克隆的乙肝病毒ｐｒｅＣ／Ｃ基因在杆状病毒载体系统中得到正确表达和加工，加工产物能够有效分泌。
The fragment of HBVpreC / C with a length of 640 bp was obtained by PCR and cloned into the vector pSXIVIV + X3 / 4 to construct the recombinant plasmid pSXIVVI + X3 / 4-pC / C. The recombinant strain TnNPV-pC / C-OCC + was constructed by co-transfection, which can form polyhedrosis and preC / C genes. The preC / C gene in this recombinant strain is doubly regulated by a tandem double promoter-synthesis promoter and a Hindlll linker-containing XIV promoter. Spodoptera frugiperda (Sf) cells infected with the recombinant strain and their culture supernatant were detected by solid phase radioimmunoassay with HBeAg and HBcAg. The infected cells and their culture supernatants showed HBeAg-positive and HBcAg-negative, respectively. Western analysis showed that the total protein of infected cells in 26kD and culture supernatant 15kD, showed a specific reaction with HBeAg monoclonal antibody bands. The results showed that the cloned hepatitis B virus preC / C gene was correctly expressed and processed in the baculovirus vector system, and the processed product can be effectively secreted.