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一步法扩增克隆IBDV上海超强毒VP2-4-3基因
Amplification and Clone of VP2-4-3 Gene of Very Virulent Infectious Bursal disease virus
【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A片段前体融合蛋白基因即VP2 4 3基因 ,将其克隆入 pGEM T载体 ,并进行序列分析 ,其与超强毒株HK4 6的核苷酸序列的同源性达 98% ,整个基因有 5个氨基酸差异 ,同源性达99.5 1% (10 0 7/ 10 12 )。
【Abstract】 The methods of reverse transcription, polymerase chain reaction (PCR) amplification, and cloning of full length VP2 4 3 gene 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 3060 base pairs (bp) of VP2 4 3 were amplified by long and accurate PCR in a single step, successfully cloned and sequenced revealing their identity of IBDV.
【Key words】 Very virulent infectious \%Bursal disease virus\%(vvIBDV); Full length cDNA; VP2 4 3 gene; Clone and sequence analysis;
- 【文献出处】 中国病毒学 ,Virologica Sinica , 编辑部邮箱 ,2002年04期
- 【分类号】S852.65
- 【被引频次】1
- 【下载频次】55