【摘要】 分离、纯化了鸡传染性法氏囊病病毒超强毒(vvIBDV)上海株(SH95)的病毒核酸dsRNA,应用随机引物将RNA反转录成cDNA,以此为模板一步扩增出全长基因组B片段,将其克隆入pGEM-T载体,并进行序列分析。克隆的B片段全长2827个核苷酸,其编码的氨基酸与超强毒株HK46的同源性达98 18%(863/879)。分子系统进化树分析表明,SH95超强毒株与美国变异株E的亲缘关系最近,但与基于基因组A片段的系统进化分析完全不同,表明该毒株在发生过程中基因重配比基因重组发挥了更重要的作用。通过对14株IBDV编码氨基酸序列的分析、比较,推测B片段上11个独特的氨基酸位点可能与毒力相关。更多还原

【Abstract】 The methods of reverse transcription,polymerase chain reaction(PCR)amplification,and cloning of full length B segment of a very virulent infectious bursal disease virus(vvIBDV)strain SH95 were developedThe use of random primer and a reverse transcriptase lacking RNase-H activity produced full length coding region and non-coding region cDNA copies of the viral genomic segmentsThe 2845 base-pairs(bp)of B segment were amplified by long and accurate PCR in a single step,successfully cloned and sequenced revealing their identity of IBDV更多还原