Species variation within the internal transcribed spacer (ITS) region of Gyrodactylus (Monogenea: Gyrodactylidae) ribosomal RNA genes
Species variation within the internal transcribed spacer (ITS) region of Gyrodactylus (Monogenea: Gyrodactylidae) ribosomal RNA genes
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DOI：
10.2307/3284442
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发表时间：
1997-04-01
影响因子：
1.3
通讯作者：
Cunningham, CO
中科院分区：
文献类型：
--
作者：
Cunningham, CO
用聚合酶链反应（PCR）扩增了三代龙属（Gyrodactylus）个体的核糖体内转录间隔区（ITS）。该反应使用与小亚基核糖体RNA基因的3'末端和大亚基核糖体RNA基因的5'末端杂交的引物扩增核糖体RNA基因簇的整个ITS 1 -5. 8 S-ITS 2区域。对野生三代虫和百里香三代虫的PCR产物进行了克隆和测序。对三代虫5.8S基因进行序列比对，确定了该基因的序列。salaris序列和血吸虫5.8S基因序列。ITS区由G. safaris、黑腹隐翅虫G.用限制性内切酶对三代虫（Gyrodactylussafaris）、德氏三代虫（Gyrodactylusderjavini）和杜氏三代虫（Gyrodactylustruttae）进行了比较分析，发现了三代虫种间限制性内切酶片段长度多态性。从序列数据中确认胸腺限制性片段大小。ITS扩增后Sau 3AI酶切可快速、清晰地区分G. safaris、黑腹隐翅虫G. derjavini和G. truttae。
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 ene 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 safaris 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. safaris, G. thymalli, Gyrodactylus derjavini, and Gyrodactylus truttae were compared by restriction enzyme analysis and interspecific restriction fragment length polymorphisms were found. Gyrodactylus safaris and G. thymalli restriction fragment sizes were confirmed from sequence data. ITS amplification followed by Sau3AI digestion enables rapid and clear differentiation of G. safaris, G. derjavini, and G. truttae.