暗纹东方鲀线粒体12S rRNA基因的克隆及序列分析

Cloning and Sequence Analysis of 12S rRNA Genes of Mitochondrial Genome from Takifugu fasciatus

利用提取纯化的暗纹东方鲀肝脏的mtDNA为模板,按照红鳍东方鲀mtDNA序列设计合成特异引物进行PCR扩增,首次克隆并测定了暗纹东方鲀mtDNA的12S rRNA基因。同时运用DNA分析软件,对暗纹东方鲀与GenBank中选取的几种同目鱼类的相应序列进行比较分析,显示暗纹东方鲀与这些鱼类的12S rRNA基因具有较高的同源性。利用分子生物学相关软件,对暗纹东方鲀及与其同目的5种鱼的12S rRNA基因序列建立NJ和MP分子进化树,并与传统的分类地位进行比较,结果表明与传统的分类地位基本吻合。

Mitochondrial eral mitochondrial genes, DNA （mtDNA） of Takifugufasciatus is prepared as templates from the liver tissue. Sevsuch as 12S rRNA and its connected tRNA genes are amplified by PCR with a pair of specific primers, which is designed based on Takifugu rubripes from Genbank. The specific PCR product are then cloned and sequenced. The total sequence has been submitted to GenBank. The 12S rRNA mtDNA of T. fasciatus is proved to have good homology with 6 species of fish which were selected from the NCBI and their sequences are recorded in the GenBank. The Cyprinus carpio is designated as outgroup. The phylogenetic trees based on the homology of the mitochondrial 12S rRNA showed that the phylogenetic relationships of T. fasciatus with other 5 kinds of fish are consistent with their morphological taxonomy.