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鸡传染性支气管炎病毒N基因片段的RT-PCR扩增及限制性内切酶分析
孙 涛1, 王 欣1, 陆 苹1
上海交通大学 农学院
摘要:
根据鸡传染性支气管炎病毒(IBV)的已报道基因序列设计了1对可扩增N基因片段的引物,并得到了预期的438 bp扩增产物;将目的条带纯化并回收后,克隆到PMD18-T质粒载体中,对插入片段进行序列测定,并与已发表的IBVN基因序列进行了比较,证实扩增和克隆的产物是正确的;根据扩增的N基因的序列,选择了BstX I酶对IBV进行分析,发现M41和澳大利亚T株BstX I酶切图谱存在明显的差异,从而建立了一种新的鉴定IBV病毒的方法。
关键词:  鸡传染性支气管炎病毒(IBV)  N基因  RT-PCR
DOI:
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基金项目:上海市教委基金资助项目(97E04)
The RT-PCR amplification and restriction endonuclease analysis of IBV N gene fragment
Abstract:
We designed a pair of primers to amplify the N gene fragment of IBR M41 and T strain based on the reported gene sequences.The anticipated 438 bp products were acquired.After purification and recovery,the products were recombined into PMD18-T vector and sequenced.The sequence were compared with the reported N gene sequences and testified that the RT-PCR products were correct.According to the sequences,we found some restriction endonuclease sites.At last,we selected BstⅪ enzyme to analyze the two IBV strains and found the maps of the two strains were significant different.So we can use this method (RFLP) to identify and distinguish IBV M41 and T strain.
Key words:  IBV  N gene  RT-PCR