| 卡氏住白细胞虫rDNA ITS-2的POR扩增与测序 |
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| 引用本文： | 李国清,薛红,朱兴全.卡氏住白细胞虫rDNA ITS-2的POR扩增与测序[J].中国农业科学,2003,36(5):573-576. |
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| 作者姓名： | 李国清 薛红 朱兴全 |
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| 作者单位： | [1]华南农业大学兽医学院，广州510642 [2]澳大利亚墨尔本大学．维多利亚3030 |
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| 基金项目： | 国家自然科学基金资助项目 ( 3 9870 5 49)，广东省自然科学基金资助项目 ( 980 13 4) |
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| 摘 要： | 运用PCR方法扩增了卡氏住白细胞虫rDNA的ITS-2及部分5．8S和28S序列(ITS2 )，并对该序列进行了克隆和测序。采用巨型裂殖体为材料，并根据Saccharomyces cerevisiae rDNA中的5．8S、28S保守序列所设计的通用引物ITS3、ITS4，进行了虫体ITS2 的PCR扩增，将所扩增片段纯化后成功地克隆于pGEM-T easy和PMDl8-T载体的T-T窗口。经双向自动测序显示，该ITS2 片段的大小为270bp；经NCBI Blast联机检索，结果显示，所扩片段中5．8S序列与S．cerevisiae的5．8S序列有部分相同，而ITS-2序列为虫体所特有；同时经wDNA-SIS软件分析，显示该ITS-2序列与S．cerevisiae间同源性相对较远，而与柔嫩艾美耳球虫间同源性相对较近，故而本试验中所测序列为卡氏住白细胞虫的ITS-2特有序列。
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| 关 键 词： | 卡氏住白细胞虫 rDNAITS-2 POR扩增 测序技术 鸡 寄生性原虫病 |
| 修稿时间： | 2002年1月17日 |
PCR Amplification and Sequencing of ITS-2 rDNA ofLeucocytozoon caulleryi |
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| LI Guo qing ,XUE Hong ,ZHU Xing quan.PCR Amplification and Sequencing of ITS-2 rDNA ofLeucocytozoon caulleryi[J].Scientia Agricultura Sinica,2003,36(5):573-576. |
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| Authors: | LI Guo qing XUE Hong ZHU Xing quan |
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| Institution: | LI Guo qing 1,XUE Hong 1,ZHU Xing quan 2 |
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| Abstract: | ITS2+ gene of Leucocytozoon caulleryi was amplified by PCR using a pair of conservative primers and cloned into the T T windows of plasmid pGEM T easy and PMD18 T carrier. The inserts were successfully sequenced and the results revealed that the ITS2+ gene was composed of 270 nucleotides while the ITS 2 gene was species specific with 113 nucleotides. The ITS 2 of L.caulleryi genes were sorted by NCBI Blast, and the degree of homology of the ITS 2 gene of L.caulleryi was compared with Eimeria tenella, Candida tropicalis, Saccharomyces kluyveri and others and analyzed by wDNASIS software. The results showed that the ITS 2 of L.caulleryi is characteristic and has the greatest similarity with E.tenella (21.2%). |
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| Keywords: | Leucocytozoon caulleryi ITS 2 PCR Cloning Sequencing |
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