眼镜王蛇线粒体基因组全序列分析Sequencingandanalysis-生物通.PDF

HEREDITAS (Beijing) ISSN 0253-9772 眼镜王蛇线粒体基因组全序列分析 陈念，赖小平 广州中医药大学中药学院，广州 510006 摘要：参照近缘物种线粒体基因序列共设计和合成了 8 对引物，结合 Ex Taq-PCR、TA 克隆和步移测序技术，文章首次获得 眼镜王蛇线粒体基因组全序列（GenBank 登录号：EU_921899 ）。该基因组全长 17 267 bp，共编码 13 个蛋白、2 个 rRNA 、 23 个 tRNA——其中 tRNA-Ile 基因发生了复制，属于一种新的蛇类物种线粒体基因排列模式，另外还含有 2 个非编码的富 含 AT 的控制区。除了 8 个 tRNA 基因和 1 个蛋白基因由 L 链编码外，其余均由 H 链编码，其中 H 链编码基因的 A 和 T 含 量接近，而 L 链上 A 的含量则明显高于T 。基于21 种蛇合并的“12S ＋16S”rRNA 基因序列的系统发育分析表明，眼镜王蛇 属与眼镜蛇属亲缘关系较近，两者与环蛇属共同构成一个单系群。作为国内外眼镜王蛇线粒体基因组全序列的首次报道， 上述结果对于蛇类物种分子系统发育和进化研究具有重要意义。 关键词：眼镜王蛇；线粒体基因组；系统发育 Sequencing and analysis of the complete mitochondrial genome of the King Cobra, Ophiophagus hannah (Serpents ： Elapidae) CHEN Nian, LAI Xiao-Ping School of Chinese Materia Medica, Guangzhou University of Chinese Medicine, Guangzhou 510006, China Abstract: We obtained the complete mitochondrial genome of King Cobra （GenBank accession number ：EU_921899 ） by Ex Taq-PCR, TA-cloning and primer-walking methods. This genome is very similar to other vertebrate, which is 17 267 bp in length and encodes 38 genes (including 13 protein-coding, 2 ribosomal RNA and 23 transfer RNA genes) and two long non-coding regions. The duplication of tRNA-Ile gene forms a new mitochondrial gene rearrangement model. Eight tRNA genes and one protein genes were transcribed from L strand, and the other genes were transcribed genes from H strand. Genes on the H strand show a fairly similar content of Adenosine and Thymine respectively, whereas those on the L strand have higher proportion of A than T. Combined rDNA sequence data (12S+16S rRNA) were used to reconstruct the phylogeny of 21 snake species for which complete mitochondrial genome sequences were available in the public databases. This large data set and an appropriate range of outgroup taxa demonstrated that Elapidae is more closely related to colubridae than viperidae, which supports the traditional