海南产花鳗鲡细胞色素b基因的克隆及序列分析

线粒体细胞色素b基因是目前研究鱼类分子系统进化的重要分子标记.笔者以日本鳗鲡线粒体基因组序列为模板设计并合成引物进行PCR扩增,并将扩增产物经琼脂糖凝胶电泳和纯化后,克隆到pMD18-T载体、测序,成功得到全长为1 140 bp的海南产花鳗鲡细胞色素b(Cyt b)基因序列.将该基因全长序列递交到Genebank数据库,获得序列登录号为EF690363.用DNA分析软件MEGA2比较了海南产花鳗鲡与递交到GenBank中的8种分布在中国东南沿海、日本海域及南太平洋海域的鳗鲡属(Anguillia)细胞色素b基因的地理差异,并构建了系统进化树.结果显示:日本产和夏威夷产花鳗的地理差异小于海南产花鳗与它们之间的地理差异.

Cloning and Sequence Analyses of Cytochrome b Gene of Anguilla marmorata from Hainan

The mitochondrial DNA cytochrome b gene is an important marker for molecular evolution study.In this study,we successfully amplified the cytochrome b gene of Anguilla marmorata from Hainan with the primers designed by referring to the sequences of Anguilla japonica.The amplification products were cloned into PMD18T-vector(TAKARA) and sequenced.The result showed that the cytochrome b gene had 1140 base pairs.The full length sequence of this gene was submitted to GenBank and obtained an Accession No.with EF690363. We compared the sequence of cytochrome b gene of Anguilla marmorata with those of other 8 Anguillas distributed in areas of Southeast China Sea,Japanese sea,and the South Pacific Ocean.We submitted sequences of cytochrome b gene into GenBank Molecular Phylogenetic tree based on these sequences to reconstruct with Neighbor-Joining method by means of MEGA2 software.The results show Hainan Anguilla marmorata has high genetic distance to others.