弓形虫ITS及5.8S序列的PCR扩增、克隆及分析

畜牧兽医学报 ›› 2005, Vol. 36 ›› Issue (1): 70-73.doi:

弓形虫ITS及5.8S序列的PCR扩增、克隆及分析

翁亚彪;谢德华;林瑞庆;李华文;张德林;吴绍强;朱兴全

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

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2005-01-25 发布日期:2005-01-25

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

1.College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642; 2.College of Basic Medical, Sun YatSen University, Guangzhou 510080; 3.Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046，China

Received:1900-01-01 Revised:1900-01-01 Online:2005-01-25 Published:2005-01-25

摘要/Abstract

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5.8S DNA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。

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.

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.

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.

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