摘要
澳大利亚T株(IBVT-)是鸡肾型传染性支气管炎病毒(IBV)主要代表毒株之一。目前,在相关生物数据库中IBV-基因序列极少。实验通过对GenBank中已有IBV基因序列比较,在IBV编码蛋白S、N、M和RdRp基因的高度同源区设计了四对引物,并通过反转录聚合酶链式反应R(TP-CR)获得特异性扩增产物。克隆测序结果证明扩增产物确为IBV相应基因片段。所获序列同IBV其它类型相关基因序列比较结果显示,编码蛋白S、N、M和RdRp基因的同源性分别为80.51%～87.22%%,85.83%～91.25%,89.63%～91.01%和85.97%～92.37%。实验丰富了生物数据库中IBVT-基因序列。所设计的引物表现很好的扩增效率和特异性,对于IBV其它毒株的检测和鉴定有一定的借鉴作用。
Avian infectious bronchitis virus Australia T（IBV-T）strain is one of the most representative strain in many different serotypes of nephropathogenic IBV strain. There is almost no its gene sequences on relative biology databanks. Four pairs of primers were designed according to reported IBV gene sequences in GenBank. The spike （S） glycoprotein, the membrane （M）glycoprotein, the nucleocapsid （N） phosphoprotein and RNA-dependent RNA polymerases（RdRp） gene were amplified by reverse transcription polymerase chain reacfion（RT-PCR） from viral RNAs of IBV-T strain respectively. The amplified products were validated by cloning and sequencing. These sequences of products were compared with other five IBV strains, The further analysis showed the homology of nucleotide sequence was 80.51%-87.22%,85.83% -91.25%,89.63% -91.01%, 85.97% -92.37% forS, N,Mand RdRp genes respectively. These primers were high efficiency and specific and may be applied to detection and identification of other IBV strain.
出处
《上海交通大学学报（农业科学版）》
2005年第3期271-274,288,共5页
Journal of Shanghai Jiaotong University(Agricultural Science)
基金
国家重点基础研究规划项目(2003CB514129)
上海市科学技术委员会技术标准专项(02DZ05030)
