【摘 要】
:
以传染性法氏囊病病毒(IBDV)浙江分离株(ZJ2000)基因组RNA为模板,采用Long-accurate RT-PCR(LA-PCR)一步法扩增并克隆了IBDV ZJ2000株基因组A节段全长cDNA.序列测定结果表明,
【机 构】
:
浙江大学动物预防医学研究所,浙江大学生物医学工程研究所
【基金项目】
:
国家高技术研究发展计划(863计划);
论文部分内容阅读
以传染性法氏囊病病毒(IBDV)浙江分离株(ZJ2000)基因组RNA为模板,采用Long-accurate RT-PCR(LA-PCR)一步法扩增并克隆了IBDV ZJ2000株基因组A节段全长cDNA.序列测定结果表明,克隆的A节段全长共3 259个核苷酸,包括5、3端的非编码区(NCRs)和2个部分重叠的开放阅读框(ORF1和ORF2),与参比的血清I型毒株核苷酸序列的同源性高达95.2%~99.2%.二级结构预测表明,在5-NCRs和3-NCRs存在1个大型的茎环发夹结构.ORF2编码145个氨基酸的VP5,与参比毒株的同源性高达98.6%~100%.ORF1编码1 012个氨基酸的VP2/VP4/VP3,在氨基酸水平上VP2、VP3、VP4与参比毒株的同源性分别达94.9%~98.8%、96.1%~98.5%、97.1%~99.2%.ZJ2000共有14~38个氨基酸的替代,其中特有氨基酸6个,变异大多数集中在VP2高变区,突变率达2.9%~11%.第2个小亲水区内280位氨基酸由S替代了N、290位M替代了L.这2个突变可能与IBDV的抗原性有关.VP2-VP4剪切位点附近511和540位氨基酸的变异可能使ZJ2000株的毒力增强.分子系统进化树分析表明,ZJ2000与欧洲Cu-1株、P2株、CEF94株和中国Harbin株的关系最近,而与欧洲、香港、日本的超强毒株和美国的变异株相对较远.
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