柞蚕微孢子核糖体基因和转录间隔区的序列及系统进化分析

The Sequence and Phylogenetic Analysis of the Ribosomal RNA Gene and the ITS Region of Nosema antheraeae

目的研究柞蚕微孢子(Nosemaantheraeae)的核糖体基因,转录间隔区(ITS)以及它们的排列顺序,为柞蚕微孢子的分类和系统进化分析提供分子生物学方面的依据。方法采用特异引物进行PCR扩增,克隆和测序。用DNAStar软件,用ClustalW的比对方法得到遗传距离和系统进化树。结果得到柞蚕微孢子的核糖体小亚基rRNA(SSUrRNA),转录间隔区(ITS),5SrRNA的全长,得到核糖体大亚基rRNA(LSUrRNA)的部分序列。结论柞蚕微孢子的核糖体基因排列顺序为LSU-ITS1-SSU-ITS2-5S与Nosemabombycis相同,是一种比较特殊的排列方式。将微孢子SSUrRNA基因和ITS结合起来分析微孢子的亲缘关系是一个新的尝试。

[Objective] This paper examines the sequence and organization of the ribosomal RNA gene, the ITS region of Nosema antheraeae, and present molecular biological evidence of Nosema antheraeae taxonomy. [Method ] To obtain the phlogenetic tree and sequence distance, specific primers were used for PCR amplification, then clone and nucleotide sequences were aligned using Clustal W ofDNAStar sotiware. [Result] The outcome entailed a full small subunit rRNAs （SSU rRNA） gene, the internal transcribed spacer （ITS）, 5S rRNA gene and part of the large subunit rRNAs （LSU rRNA） gene of Nosema antheraeae. [Conclusion] The organization of the ribosomal RNA gene of Nosema antheraeae follows a special organizational pattern： LSU-ITS1-SSU-ITS2-5S. Elucidating phylogenetic relationships of microsporidia species by combining information from SSU rRNA and ITS sequence is a new experiment.