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目的 克隆和测定剑尾鱼 (Xiphophorushelleri)线粒体细胞色素b基因 (cytb)的全序列。方法 提取剑尾鱼肝脏的总DNA。设计合成特异引物进行PCR扩增。扩增产物经琼脂糖电泳检测、纯化后克隆到pGEM Teasyvectorsystem中的T载体上 ,筛选转化子 ,提取质粒 ,酶切鉴定。挑取重组质粒pGEM T xhcytb 11进行序列测定。结果 获得了剑尾鱼线粒体cytb基因的全序列 ,共 114 0bp。结论 用BLAST与GenBank中的线粒体DNA序列进行比较 ,显示剑尾鱼与其他鱼类的cytb基因具有较高的同源性 ;根据剑尾鱼与其他 13种鱼的cytb基因序列同源性所建立的进化树 ,与传统的分类地位基本吻合
Objective To clone and determine the complete sequence of mitochondrial cytochrome b gene in Xiphophorus helleri. Methods The total DNA of the swordtail fish liver was extracted. Design and synthesis of specific primers for PCR amplification. The amplified product was detected by agarose gel electrophoresis, purified and cloned into the T vector in pGEM Teasyvectorsystem. The transformants were screened and the plasmids were extracted and identified by restriction enzyme digestion. The recombinant plasmid pGEM T xhcytb 11 was picked for sequencing. Results The full-length sequence of mitochondrial cytb gene was obtained from a total of 114 0bp. Conclusion The comparison between mitochondrial DNA sequences of BLAST and GenBank showed that Swordfish has high homology with other cytb genes. Based on the sequence homology of cytb gene between Swordfish and other 13 species of fish, Phylogenetic tree, basically consistent with the traditional classification status