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PCR介导HaNPV罹病幼虫潜伏期cDNA文库的构建

Construction of Helicoverpa armigera Nuclear Polyhedrovirus cDNA Library During Latent Periods by Polymerase Chain Reaction

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【作者】 白志强张友清

【Author】 BAI Zhi qiang, ZHANG You qing (Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071,China)

【机构】 中国科学院武汉病毒研究所!武汉430071

【摘要】 以罹病棉铃虫幼虫为材料提取总RNA ,反转录合成cDNA第一链 ,加oligo(dG)同聚尾 ,PCR扩增合成双链cDNA ,克隆到 pGEM T质粒载体中。随机筛选文库中阳性克隆 ,经酶切分析 ,cDNA插入片段大小在 0 .3~ 1.1kb之间。文库中原代重组子数为 1.66× 10 5,重组百分比为 87.8%。重组质粒的杂交分析表明 ,文库中HaNPV基因的cDNA克隆所占比例超过 50 %。

【Abstract】 A general cDNA library which was from infected larvae dueing the latent periods has been constructed. The first cDNA strand was synthesized from total RNA by M MLV transcriptase. After oligo(dG) tailing the cDNA was amplified by polymerase chain reaction. The double strands cDNA were ligated to pGEM T easy vectors. Screeing of cDNA library showed that the length of inserts was from 0.3 to 1.1 kilobases. The capacity of cDNA library was 1.66×10 5 clones. Moreover, the number of cDNA clones of HaNPV was more than 50%.

  • 【文献出处】 中国病毒学 ,Virologica Sinica , 编辑部邮箱 ,2000年04期
  • 【分类号】Q78
  • 【被引频次】2
  • 【下载频次】19
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