传染性法氏囊病病毒疫苗株B87基因组A节段的克隆和序列分析  

Cloning and Sequencing of Genomic A Segment of Infectious Bursal Disease Virus Vaccine Strain B87

作　　者：刘锴[1] 王兴龙[1] 王学理[2] 任林柱[3] 闫广谋[3] 段小宇[3] 

机构地区：[1]军事医学科学院十一所 [2]内蒙古民族大学动物科技学院,通辽028000 [3]吉林大学畜牧兽医学院,长春130062

出　　处：《中国生物制品学杂志》2007年第9期646-651,共6页Chinese Journal of Biologicals

基　　金：军事医学科学院科技创新基金项目;吉林省科技厅科技发展计划项目(20050549)

摘　　要：目的 获得传染性法氏囊病病毒疫苗株B87 A节段全长cDNA,并对其进行序列分析.方法 使用蛋白酶K、酚/氯仿抽提法提取病毒基因组,用LiCl分级沉淀方法纯化病毒基因组dsRNA,通过RT-PCR一步扩增获得A节段的全长cDNA.将其克隆至pGEM-T载体上,测序,并用DNAStar软件进行序列分析.结果 测序结果表明,克隆的B87株A节段全长为3 260 bp,与Gt株的同源性最高为99.5%,与其他毒株的核苷酸同源性介于95.2%～99.4%之间,与Ⅱ型毒株OH仅为84.0%.结论 通过对20株IBDV编码氨基酸序列的分析、比较,推测A片段上7个独特的氨基酸位点可能与毒力相关.Objective To clone and sequence the full-length cDNA of genomic segment A of infectious bursal disease virus （IBDV） vaccine strain B87. Methods Extract the genomic RNA of IBDV by protease K digestion and phenol/chloroform extraction, then purify the genomic dsRNA by LiCl fractional precipitation for amplification of full-length cDNA of genomic segment A of IBDV strain 1387 by one-step RT-PCR. Insert the amplified cDNA into pGEM-T vector,then sequence and analyze the result by using DNAStar software. Results Nucleotide sequencing proved that the homologies of cloned genomic A segment of IBDV vaccine strain B87, at a fuU-length of 3260 bp,were 99. 5% and 95. 2% -99.4% respectively to those of Gt and other strains,but was only 84. 0% to that of strain OH of IBDV type I1. Conclusion The analysis of deduced amino acid sequences of 20 IBDV strains indicated that the 7 characteristic amino acid sites of A segment might be related to virulence.

关 键 词：传染性法氏囊病病毒 全长CDNA A节段 克隆 序列分析 

分 类 号：S858.3[农业科学—临床兽医学] Q78[农业科学—兽医学]