直额裸腹溞线粒体DNA COⅠ基因的克隆及序列分析

Cloning and sequencing analysis of COⅠ in water fleas Moina rectirostris

为研究直额裸腹溞Moina rectirostris线粒体细胞色素氧化酶Ⅰ亚基(COⅠ)基因的特点,使用WizardTM基因组DNA纯化试剂盒提取直额裸腹溞DNA,以其DNA为模板,用线粒体COⅠ基因通用引物进行PCR扩增、测序。结果表明:直额裸腹溞线粒体DNA COⅠ基因部分cDNA序列长度为709 bp,其中碱基A、G、T、C含量分别为26.23%、19.89%、38.50%、15.37%,A+T含量(64.74%)明显高于G+C含量(35.26%);将该基因的cDNA序列与裸腹溞科5个种的同源序列进行比对,并用邻接法(NJ)构建系统进化树,结果显示,直额裸腹溞与短型裸腹溞的遗传距离最小,同属于一个分支,亲缘关系最近。研究表明,应用COⅠ基因序列可以有效地区别裸腹溞科各个种类,与传统形态学分类学的结果基本一致。

DNA was extracted and partial COⅠ was sequenced from water fleas Moina rectirostris using WizardTM genomicDNA purification kit,and the extracted DNA was used as a template in order to study the characteristics of mitochondrial cytochrome c oxidase subunit 1（COⅠ） in the water fleas.The sequence analysis revealed that the sequences had length of 709 bp,with 26.23% in A,19.89% in G,38.50% in T and 15.37% in C,and,higher in A ＋ T（64.74%） than in G ＋ C（35.26%）.The phylogenetic tree was constructed with neighbor-joining of the six species by five species in Moinidae from the Gen Bank and the alignmented sequences.The results showed that the genetic distance between Moina rectirostris and Moina brachiata was the shortest,which belonged to a branch and the closest relationship.The genetic distance and NJ phylogenetic tree indicated that the species in Moinidae could be effectively separated,which was consistent with that by traditional morphological taxonomy.