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三代虫(单殖亚纲:三代虫科)核糖体RNA基因内部转录间隔区(ITS)的种间变异

Species variation within the internal transcribed spacer (ITS) region of Gyrodactylus (Monogenea:Gyrodactylidae) ribosomal RNA genes.

作者信息

Cunningham C O

机构信息

SOAEFD Marine Laboratory, Aberdeen, United Kingdom.

出版信息

J Parasitol. 1997 Apr;83(2):215-9.

PMID:9105299
Abstract

The ribosomal internal transcribed spacer (ITS) region from individual Gyrodactylus specimens was amplified by polymerase chain reaction (PCR). The reaction amplified the entire ITS1-5.8S-ITS2 region of the ribosomal RNA gene cluster using primers that hybridize to the 3' terminus of the small subunit and the 5' terminus of the large subunit ribosomal RNA genes. The PCR products from Gyrodactylus salaris and Gyrodactylus thymalli were cloned and sequenced. The Gyrodactylus 5.8S gene was identified following comparative alignment of the G. salaris sequence and a Schistosoma 5.8S gene sequence. The ITS regions from G. salaris, G. thymalli, Gyrodactylus derjavini, and Gyrodactylus truttae were compared by restriction enzyme analysis and interspecific restriction fragment length polymorphisms were found. Gyrodactylus salaris and G. thymalli restriction fragment sizes were confirmed from sequence data. ITS amplification followed by Sau3AI digestion enables rapid and clear differentiation of G. salaris, G. derjavini, and G. truttae.

摘要

通过聚合酶链反应(PCR)扩增单个三代虫标本的核糖体内部转录间隔区(ITS)。该反应使用与小亚基3'末端和大亚基核糖体RNA基因5'末端杂交的引物,扩增核糖体RNA基因簇的整个ITS1-5.8S-ITS2区域。对来自鲑三代虫和百里香三代虫的PCR产物进行克隆和测序。通过将鲑三代虫序列与血吸虫5.8S基因序列进行比较比对,鉴定出三代虫5.8S基因。通过限制性酶切分析比较了鲑三代虫、百里香三代虫、德氏三代虫和鳟三代虫的ITS区域,并发现了种间限制性片段长度多态性。从序列数据中确认了鲑三代虫和百里香三代虫的限制性片段大小。ITS扩增后用Sau3AI消化能够快速、清晰地区分鲑三代虫、德氏三代虫和鳟三代虫。

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