乙型肝炎病毒表面抗原基因的克隆及序列分析  

Cloning and sequencing of human hepatitis B virus S gene

作　　者：李小月[1] 何金生[2] 沈继龙[1] 汪学龙[1] 姜山[2] 

机构地区：[1]安徽医科大学病原生物学教研室,合肥230032 [2]安徽医科大学免疫学教研室,合肥230032

出　　处：《热带病与寄生虫学》2004年第2期75-76,94,共3页Journal of Tropical Diseases and Parasitology

基　　金：国家自然科学基金(30371320);安徽省自然科学基金(03043601);安徽省教育厅自然科学研究项目(2003kj182)

摘　　要：目的克隆并鉴定乙型肝炎病毒表面抗原基因,为下一步构建该重组腺病毒载体以及进一步研究腺病毒载体的包装及在乙型肝炎病毒基因治疗中的作用。方法参照人 HBV adr 亚型序列,设计和合成 S 基因特异引物。应用 PCR 技术,从含有 HBsAg 的 HBV DNA 中扩增目的 DNA,通过 TA 连接将其克隆人 pGEM-T easy 载体,经限制性内切酶 BglⅡ/SalⅠ鉴定后,进一步测序鉴定。结果从乙肝表面抗原阳性(HBsAg+)血清中成功提取 HBV DNA,并以此 DNA 为模板,成功扩增出 S 基因,测序结果与 GenBank 中注册的相应序列比对,核苷酸序列的同源性高达92％～99％,预测氨基酸序列同源性亦达82％。结论从 HBsAg 阳性血清中成功克隆出 S 基因序列,为进一步构建重组腺病毒及后续实验奠定了基础。Objective To clone and identify S gene of human hepatitis B virus surface antigen for ap- proach to construction of its recombinant adenovirus vector and for studying the package of HBsAg-expressing recombinant adenovirus vector and its effection on genetic therapy of human hepatitis B virus.Methods A pair of primers were designed based on the subtype adr DNA sequence of hepatitis B virus S-gene.Speific fragment of S gene was obtained by PCR amplification from HBV positive serum.The PCR products were lig- ated into pGEM T easy vectors.The PCR-generated DNA fragments were sequenced following digestion of Bgl Ⅱ and Sal Ⅰ.Results The clone DNA fragments possess 92-99% homology with the published HBV DNA sequence.There is 82% of identity in deduced amino acid sequence.Conclusion DNA fragments of HBV S-gene were cloned and sequenced,which having 92-99% homology with the published HBV DNA sequence.

关 键 词：表面抗原基因 乙型肝炎病毒 克隆 序列分析 乙肝表面抗原阳性 重组腺病毒载体 HBsAg 病毒基因治疗 限制性内切酶 HBVDNA ADR亚型 PCR技术 核苷酸序列 序列同源性 S基因序列 特异引物 序列比对 阳性血清 鉴定 步研究 氨基酸 

分 类 号：R373.21[医药卫生—病原生物学] R512.62[医药卫生—基础医学]