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弓形虫ITS及5.8S序列的PCR扩增、克隆及分析
引用本文:翁亚彪,谢德华,林瑞庆,李华文,张德林,吴绍强,朱兴全.弓形虫ITS及5.8S序列的PCR扩增、克隆及分析[J].畜牧兽医学报,2005,36(1):70-73.
作者姓名:翁亚彪  谢德华  林瑞庆  李华文  张德林  吴绍强  朱兴全
作者单位:1. 华南农业大学兽医学院,广州,510642
2. 中山大学基础医学院,广州,510080
3. 中国农业科学院兰州兽医研究所,兰州,730046
基金项目:国家杰出青年科学基金项目(30225033),广东省科技攻关计划项目(2004B20201008)
摘    要:通过对国内来源于不同宿主的ZS人株、SH人株、CN猪株、QH绵羊株4个弓形虫虫株,以及国际标准强毒株RH株的核糖体DNA内转录间隔区(ITS)及5.8SDNA序列进行PCR扩增、克隆、测序和序列分析,旨在对国内不同宿主间弓形虫虫株的遗传变异情况进行分析和验证。为分子遗传学和分子诊断学研究提供资料。结果显示:QH绵羊株、ZS人株、SH人株、CN猪株的ITS及5.8S序列完全一致。且与GenBank上注册RH株的ITS及5.8S序列也一致;仅实验室传代保存的RH株的ITS2序列与其它4株的ITS2有2个碱基的差异。结果表明ITS可作为分子标记用于弓形虫与其它原虫的种间鉴定。但不适合用于弓形虫种内遗传变异的研究。

关 键 词:弓形虫  SH  序列  RH  克隆  宿主  CN  绵羊  PCR扩增  强毒株
文章编号:0366-6964(2005)01-0070-04

PCR Amplification,Cloning and Sequencing Analysis of ITS and 5.8S of rDNA from Toxoplasma gondii
WENG Ya-Biao,XIE De-hua,LIN Rui-qing,LI Hua-wen,ZHANG De-lin,WU Shao-qiang,ZHU Xing-quan.PCR Amplification,Cloning and Sequencing Analysis of ITS and 5.8S of rDNA from Toxoplasma gondii[J].Acta Veterinaria et Zootechnica Sinica,2005,36(1):70-73.
Authors:WENG Ya-Biao  XIE De-hua  LIN Rui-qing  LI Hua-wen  ZHANG De-lin  WU Shao-qiang  ZHU Xing-quan
Abstract:The aims of this study were to analyze the genetic variation among strains of Toxoplasma gondii from different hosts and geographic origins in China, and to establish the foundation for further studies on molecular genetics and diagnostics of T.gondii. The internal transcribed spacer (ITS) and 5.8S of rDNA from ZS (human), SH (human), CN (pig), QH (sheep) and RH strains of T. gondii were amplified by PCR, cloned and sequenced. The results showed that ITS and 5.8S sequences of QH, ZS, SH, CN strains were identical, which were consistent with the sequences of the RH strain registered in GenBank; but the ITS2 sequence of the RH strain differed at two bases when compared with that of the other 4 strains. The results indicated that ITS sequences provide useful genetic markers for the identification and differentiation of T. gondii from other related protozoa, but the sequences are not suitable markers for studying the genetic variation within T. gondii.
Keywords:Toxoplasma gondii  PCR  internal transcribed spacer  5  8S  sequence analysis
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