桂林地区麻鸭DHBV基因全长克隆和序列分析以及DHBV定量检测方法的建立  

Cloning and Sequence Analysis of the DHBV Genome of the Brown Ducks in Guilin Region and Establishment of the Quantitative Method for Detecting DHBV

作　　者：苏何玲[1] 黄日东[1] 何松青[2] 徐庆[1] 朱华[1] 莫之婧[1] 刘青波[1] 刘永明[1] 

机构地区：[1]桂林医学院生物化学与分子生物学教研室,桂林541004 [2]桂林医学院附属医院,桂林541004

出　　处：《病毒学报》2013年第2期180-184,共5页Chinese Journal of Virology

基　　金：广西自然科学基金项目2012GXNSFAA053146;广西中医药科技专项课题GZKZ-Z1107;桂林科技攻关项目10124008-15;桂林医学院项目驱动研究项目JG2010056资助

摘　　要：携带鸭乙肝病毒(DHBV)麻鸭作为乙肝动物模型被广泛应用于抗HBV药物活性和毒性研究,广西桂林地区麻鸭具有较高的DHBV携带率,但其DHBV基因组结构特点尚未见报道。本研究克隆了桂林麻鸭DHBV基因组,其序列全长3 027bp,ORF finder分析发现桂林麻鸭DHBV除具有S、P、C开放阅读框架外,还有一编码未知多肽的开放阅读框架,Vector NTI 8.0分析显示该多肽具有与HLA*0201结合的模体结构。比对不同国家及地区DHBV序列,并以Mega5.0生成进化树提示DHBV序列差异无明显的地域分布特点。以所构建的DHBV S基因重组质粒pGEM-DHBV-S为标准品,建立了荧光定量PCR检测DHBV方法。本研究结果为应用桂林麻鸭作为乙肝动物模型奠定了实验基础。Brown ducks carrying DHBV were widely used as hepatitis B animal model in the research of the activity and toxicity of anti-HBV dugs. Studies showed that the ratio of DHBV carriers in the brown ducks in Guilin region was relatively high. Nevertheless, the characters of the DHBV genome of Guilin brown duck remain unknown. Here we report the cloning of the genome of Guilin brown duck DHBV and the sequence analysis of the genome. The full length of the DHBV genome of Guilin brown duck was 3 027bp. Analysis using ORF finder found that there was an ORF for an unknown peptide other than S-ORF, PORF and C-ORF in the genome of the DHBV. Vector NTI 8.0 analysis revealed that the unknown peptide contained a motif which binded to HLA ＊ 0201. Aligning with the DHBV sequences from different countries and regions indicated that there were no obvious differences of regional distribution among the sequences. A fluorescence quantitative PCR for detecting DHBV was establishment based on the recombinant plasmid pGEM-DHBV-S constructed. This study laid the groundwork for using Guilin brown duck as a hepatitis B animal model.

关 键 词：鸭乙型肝炎病毒 序列分析 荧光定量PCR 

分 类 号：R373.21[医药卫生—病原生物学]