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利用克隆的核糖体DNA片段和聚合酶链反应进行序列扩增对新描述的亚洲带绦虫进行鉴定和检测。

Characterization and detection of a newly described Asian taeniid using cloned ribosomal DNA fragments and sequence amplification by the polymerase chain reaction.

作者信息

Zarlenga D S, McManus D P, Fan P C, Cross J H

机构信息

U.S. Department of Agriculture, Livestock and Poultry Sciences Institute, Beltsville, Maryland 20705.

出版信息

Exp Parasitol. 1991 Feb;72(2):174-83. doi: 10.1016/0014-4894(91)90135-j.

Abstract

DNA isolated from a newly described taeniid from Taiwan which shows adult characters indistinguishable from those of Taenia saginata was compared to DNA from T. saginata and nine other cestodes by restriction endonuclease digestion of genomic DNA and Southern blot analysis using 32P-labeled total cestode RNA and cloned ribosomal RNA gene fragments as probes. Hybridization patterns of Taiwan Taenia DNA revealed distinct variations from that of T. saginata and Taenia solium as well as all other cestode DNAs examined; however, similarities in restriction maps and sequence data between cloned ribosomal gene fragments from Taiwan Taenia (pTTr 3.1) and T. saginata (pTSgr 3.1 and PTSgr 2.4, respectively) suggest close evolutionary relatedness between these two taeniids. DNA sequence amplification from genomic DNA using oligonucleotide primers homologous to regions on both the 2.4- and 3.1-kb fragments was able to delineate between Taiwan Taenia and T. saginata by generating 1.0- and 0.29-kb fragments, respectively. Results demonstrated that Taiwan Taenia is not exclusive to Taiwan but exists in other parts of Eastern Asia and that adult morphology is insufficient for its detection in other locations. Results further support biological data indicating that Taiwan Taenia and T. saginata, although similar morphologically, are distinct genotypes.

摘要

从台湾一种新描述的带绦虫分离出的DNA,其成虫特征与牛带绦虫难以区分,通过对基因组DNA进行限制性内切酶消化,并使用32P标记的总绦虫RNA和克隆的核糖体RNA基因片段作为探针进行Southern印迹分析,将其与牛带绦虫及其他9种绦虫的DNA进行比较。台湾带绦虫DNA的杂交模式显示出与牛带绦虫、猪带绦虫以及所有其他所检测的绦虫DNA明显不同;然而,台湾带绦虫(pTTr 3.1)与牛带绦虫(分别为pTSgr 3.1和PTSgr 2.4)克隆的核糖体基因片段之间的限制性图谱和序列数据相似性表明这两种带绦虫之间存在密切的进化关系。使用与2.4 kb和3.1 kb片段上区域同源的寡核苷酸引物从基因组DNA进行DNA序列扩增,分别产生1.0 kb和0.29 kb片段,从而能够区分台湾带绦虫和牛带绦虫。结果表明,台湾带绦虫并非仅存在于台湾,而是存在于东亚其他地区,并且成虫形态不足以在其他地区检测到它。结果进一步支持了生物学数据,表明台湾带绦虫和牛带绦虫虽然形态相似,但却是不同的基因型。

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