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水泡性口炎病毒N基因的克隆及其PCR检测方法的建立

Cloning of N gene of vesicular stomatitis virus (VSV) and establishment of the PCR method for dectection of VSV

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【作者】王海霞；郑增忍；龚振华；张彦明；宫云浩；孙淑芳；李会荣；郭福生；蒋正军；

【Author】 WANG Hai-xia~(1,2), ZHENG Zeng-ren~1, GONG Zhen-hua~2, ZHANG Yan-ming~1, GONG Yun-hao~3, SUN Shu-fang~2, LI Hui-rong~1, GUO Fu-sheng~2, JIANG Zheng-jun~2 (1. College of Animal Science and Technology, Northwest Sci-Tech University of Agriculture and Forestry, Yangling 712100,China; 2. Monitoring and Detection Center of the Health and Quality of Animals and Animal Products, Animal Quarantine Institute of Ministry of Agriculture,Qingdao 266032,China; 3. Department of Virology and Molecular Biology,Vancouver Virology Research Center, Canada)

【机构】西北农林科技大学动物科技学院；农业部动物检疫所动物及动物产品卫生质量监督检验测试中心；温哥华病毒研究中心病毒及分子生物学部；农业部动物检疫所动物及动物产品卫生质量监督检验测试中心陕西杨凌712100；山东青岛266032；陕西杨凌712100；加拿大温哥华；山东青岛266032；

【摘要】根据国外发表的水泡性口炎病毒 (VSV)基因组核苷酸序列 ,设计了 2对特异性引物 ,通过RT PCR方法得到N基因。将此基因与 pGEM TEasyVector相连 ,通过蓝白斑筛选出阳性克隆 ,碱裂解法小剂量提取质粒。经测定 ,N基因与参考序列同源性高达 99%。同时根据克隆产物进行亚克隆 ,建立了VSV的PCR检测方法。更多还原

【Abstract】 A pair of primers was designed to amplify N gene of VSV according to the published relevant sequence of the virus. The N gene was obtained by RT-PCR and sequenced after being directly inserted into the pGEM-T Easy Vector. The result showed that the homology between N gene and the published sequence was 99%.Based on the cloning of N gene,another pair of primers was designed. Through subcloning,a new PCR method had been established to detect the VSV.更多还原

【关键词】水泡性口炎病毒； N基因；克隆； PCR检测；
【Key words】 vesicular stomatitis virus； N gene； cloning； PCR detection；

【文献出处】中国兽医科技 ,Chinese Journal of Veterinary Science and Technology , 编辑部邮箱 ,2003年12期

【分类号】S852.65
【被引频次】3
【下载频次】158

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