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有齿食道口线虫ITS及5.8S DNA片段的PCR扩增、克隆及序列分析

PCR Amplification, Cloning and Sequence Analysis of the ITS and 5.8S rDNA of Oesophagostomum Isolates

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【作者】 林瑞庆; 陈丽莎; 翁亚彪; 吴绍强; 邹丰才; 李明伟; 宋慧群; 朱兴全;

【Author】 LIN Rui-qing , CHEN Li-sha , WENG Ya-biao , WU Shao-qiang , ZOU Feng-cai,LI Ming-wei, SONG Hui-qun , ZHU Xing-quan (College of Veterinary Medicine, South China Agricultural University,Guangzhou 510642)

【机构】 华南农业大学兽医学院; 华南农业大学兽医学院 广州510642; 广州510642; 广州510642;

【摘要】 运用 PCR 方法以保守引物 NC5 及 NC2 扩增了从广东阳江地区猪体分离的食道口线虫 rDNA 的内转录间隔区(ITS)及 5.8S 序列。将 PCR 扩增出的片段纯化后克隆至 pGEM-T Easy 载体,用 PCR 技术及酶切鉴定阳性菌落,对阳性菌落质粒 DNA 进行测序。结果表明,扩增的片段大小为 828 bp,包含部分的 18S、28S 及全部的 ITS-1(362 bp)、5.8S(153 bp)及 ITS-2(217 bp)序列。序列比较表明,该食道口线虫为有齿食道口线虫。本研究在国际上首次报道了中国猪有齿食道口线虫的 ITS 及 5.8S 序列,为食道口线虫的分子生物学的进一步研究奠定了基础。

【Abstract】 The internal transcribed spacer (ITS) and 5.8S rDNA of Oesophagostomum spp isolated from Guangdong Province was amplified by PCR using a pair of conserved primers and the amplicons were cloned into pGEM-T Easy vector. The inserts were successfully sequenced, and the results revealed that the inserts were 828 bp in length and consisted of partial 18S, 28S, and complete ITS-1, 5.8S and ITS-2 DNA sequences. The Oesophagostomum isolates were identified as O.dentatum based on ITS-2 sequence. It was the first time that the complete sequence of ITS-1 and 5.8S rDNA of Oesophagostomum dentatum was reported. The results of the present study have laid a foundation for further studies of Oesophagostomum.

【关键词】 食道口线虫; ITS; 5.8S; PCR; 克隆; 序列分析;
【Key words】 Oesophagostomum; ITS; 5.8S; PCR; Cloning; Sequence analysis;
【基金】 国家杰出青年科学基金资助项目(30225033);教育部优秀青年教师资助计划项目(2001056401);华南农业大学校长基金资助项目
  • 【文献出处】 中国农业科学 ,Scientia Agricultura Sinica , 编辑部邮箱 ,2005年03期
  • 【分类号】S852.7
  • 【被引频次】26
  • 【下载频次】376
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