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泛亚型口蹄疫病毒O/China/99缺失毒株全长cDNA分子克隆的构建

Construction of full-length cDNA clone of deleted foot-and-mouth disease virus Pan-Asia strain O/China/99
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摘要 设计并合成了4条O型口蹄疫病毒(FMDV)泛亚型代表毒株O/China/99基因组的引物,利用RT-PCR扩增各基因片段,酶切后连接到pOK12载体上,然后人工合成一段缺失PKs的5′UTR序列,利用两端的限制性内切酶位点,将该基因片段插入到上述载体中。酶切、PCR和序列测定结果显示,O/China/99株全基因组由8 200个核苷酸组成,构建的感染性cDNA与原毒株的核苷酸序列同源性为99.1%。结果表明,O/China/99缺失毒株全长cDNA分子克隆的构建成功。 Two pairs of primers were designed and synthesized according to the genome sequence of deleted foot-and-mouth disease virus（FMDV） type O Pan-Asia strain O/China/99. Two amplified fragments from the genome of O/China/99 strain and a artificially-synthesized 5＇UTR sequence deleted PKs were cloned into pOK12 vector. The recombinant was identified by restriction enzyme digestion, PCR and sequencing,respectively. The whole genome of FMDV O/China/99 strains was 8 200 nt in length. The nuc leotide sequence of the full length eDNA shared 99.1% identities with the strain O/China/99. The results showed that the full length eDNA clone of FMDV O/China/99 strain was constructed successfully.
作者 吕建亮 张永光 王永录 潘丽 刘力宽 方玉珍 蒋守田 张维德
机构地区 中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室农业部畜禽病毒学重点开放实验室
出处 《中国兽医科学》 CAS CSCD 北大核心 2008年第11期925-928,共4页 Chinese Veterinary Science
基金 甘肃省自然科学基金项目(2007GS04449)
关键词 口蹄疫病毒 全长CDNA 反向遗传学 PKS foot-and-mouth disease virus full length eDNA reverse genetics PKs
分类号 S852.659.6 [农业科学—基础兽医学] Q786 [生物学—分子生物学]
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参考文献9

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中国兽医科学
2008年 第11期

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