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目的:克隆HBeAg结合蛋白未知功能新基因 HBeBP4的剪切体基因HBeBP4A,构建其真核表达载体,并应用生物信息学初步探讨其结构及功能．方法:应用PCR技术从HepG2细胞提取的cDNA扩增HBeBP4基因的剪切体基因 HBeBP4A,选用pGEM-Teasy载体进行TA 克隆,通过PCR,限制性酶切分析及测序进行鉴定,再将其亚克隆到真核表达载体 pcDNATM3．1/myc-HisA,通过PCR,限制酶切分析进行鉴定,并应用生物信息学初步分析其物理化学性质,蛋白质结构和功能．结果:成功扩增出HBeBP4剪切体基因,命名为HBeBP4A．利用生物信息学分析推定其 ORF为1104个核苷酸(nt),编码产物为367个氨基酸残基(aa)．结论:发现了HBeAg结合蛋白4(HBeBP4)基因的剪切体HBeBP4A,构建了pcDNATM3．1／ myc-HisA真核表达载体．
OBJECTIVE: To clone HBeBP4A gene of HBeBP4, a novel function gene of unknown function of HBeAg binding protein, and to construct its eukaryotic expression vector. The structure and function of HBeBP4A was explored by bioinformatics. Methods: The cloned HBeBP4A gene of HBeBP4 gene was amplified from HepG2 cells by PCR and cloned into pGEM-Teasy vector. The cloned products were identified by PCR, restriction analysis and sequencing, then subcloned into The eukaryotic expression vector pcDNATM3.1 / myc-HisA was identified by restriction endonuclease analysis and bioinformatics analysis of its physico-chemical properties, protein structure and function. Results: The HBeBP4 gene was successfully amplified and named as HBeBP4A. Bioinformatics analysis suggested that the ORF was 1104 nucleotides (nt) and the encoded product was 367 amino acid residues (aa). Conclusion: The cloned HBeBP4A gene of HBeAg binding protein 4 (HBeBP4) gene was constructed and the eukaryotic expression vector pcDNATM3.1 / myc-HisA was constructed.