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中国农学通报

中国农学通报 ›› 2011, Vol. 27 ›› Issue (7): 304-308.

所属专题： 生物技术； 畜牧兽医

• 畜牧 动物医学 蚕 蜂 • 上一篇    下一篇

绵羊痘病毒RPO30基因的克隆及序列分析

赵志荀 吴国华   

  • 收稿日期:2010-09-19 修回日期:2010-10-21 出版日期:2011-04-05 发布日期:2011-04-05
  • 基金资助:

    农业部转基因生物新品种培育重大专项;甘肃省重大科技专项

Sheeppox Virus RPO30 Gene Cloning and Sequence Analysis

  • Received:2010-09-19 Revised:2010-10-21 Online:2011-04-05 Published:2011-04-05

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摘要：

选择羊痘病毒RNA聚合酶RPO30进行分析,为进行RNAi羊痘病毒研究,进行目的基因的初步筛选。PCR扩增RPO30基因,连入pMD18-T simple 载体并转化DH5a大肠杆菌。经蓝白斑筛选挑选白斑制备质粒，经双酶切、PCR及测序鉴定。结果成功克隆了RPO30基因，克隆的SSPV RPO30基因ORF为585bp,编码193个氨基酸，阅读框内有一个HindIII酶切位点，测序结果与GenBank数据库中不同痘病毒毒株之间同源性存在着明显区别。进一步经过生物学软件分析表明RPO30蛋白质氨基酸4～12、18～26、50～61、68～92，176-190位之间区域形成活性中心的可能性较大，选择这些区域进行作用可能会造成该基因产物酶的失活，从而高效抑制病毒的复制。

关键词: 植被类型, 植被类型

Abstract:

Abstract: To research the goat pox virus by RNAi, the RNA polymerase 30 gene (RPO30) as a target sequence was analyzed. In the study, RPO30 was cloned by PCR, and sequencing shows that the open reading frame is 585bp that can code 193 amino acids,and there is a HindIII enzyme cut sites located during 110 to 117bp. The RPO30 sequence we cloned has different homology between different pox virus strains in the GenBank database. More bio-informatics software analyzed show that the amino acids site 4～12aa、18～26aa、50～61aa、68～92aa and176～190aa have more possibility of forming enzyme activity center. So, we should select these sites to inference.

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