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Position: Home > Articles > Cloning and Sequence Character Analysis of Squaliobarbus curriculus Mitochondrial ND6 Gene Journal of Anhui Agricultural Sciences 2010,38 (1) 95-97

赤眼鳟线粒体NADH脱氢酶6基因克隆及序列特征分析

作  者:
沈莹;左成学;吴敏;王星果;张营;徐建荣;顾志良
单  位:
常熟理工学院生物系
关键词:
赤眼鳟;ND6基因;DNA序列;系统进化
摘  要:
[目的]根据赤眼鳟线粒体NADH脱氢酶6(ND6)基因序列分析不同种鱼类的分子进化关系。[方法]根据其他鱼类的线粒体ND6基因及侧翼序列设计引物,采用PCR扩增并克隆、测序赤眼鳟ND6基因,通过MEGA4.0 Neighbor-Joining法分析不同鱼类的进化关系。[结果]赤眼鳟(Squaliobarbus curriculus)线粒体基因组ND6基因序列长度为522 bp,编码173个氨基酸。该基因碱基组成为:A41.5%、C32.7%、G12.4%、T13.4%,其中A+T含量(54.9%)高于G+C含量(45.1%)。将赤眼鳟ND6基因序列与GenBank中其他11种鱼类ND6基因序列进行比对发现,赤眼鳟ND6核酸序列与团头鲂的同源性最高,为90.8%;与红点鲑鱼的同源性最低,为73.0%。赤眼鳟ND6氨基酸序列与其他物种ND6氨基酸进行比对发现,赤眼鳟与团头鲂同源性最高,为98.3%;与大黄鱼的同源性最低,为75.7%。以ND6氨基酸序列采用NJ法构建12种鱼类的系统发生树基本反映了它们的种间、属间和科间的关系。[结论]首次得到了赤眼鳟线粒体ND6基因序列,并确定了赤眼鳟与其他11种鱼类的分子进化关系。
译  名:
Cloning and Sequence Character Analysis of Squaliobarbus curriculus Mitochondrial ND6 Gene
作  者:
SHEN Ying et al(Department of Biology,Changshu Institute of Technology,Changshu,Jiangsu 215500)
关键词:
Squaliobarbus curriculus;ND6 gene;DNA sequence;Phylogeny
摘  要:
[Objective]The study aimed to obtain the sequence of NADH dehydrogenase 6(ND6) gene of Squaliobarbus curriculus and reconstruct the phylogeny of 12 fish species based on the ND6 sequence.[Method] The primers were designed based on ND6 gene and its flank sequence in carp and other fish species.Fragments were amplified from the genomic DNA of S.curriculus and then cloned into PMD18.The positive clones were sequenced.The phylogenetic tree of 12 fish species were reconstructed using NJ method by MEGA4.[Result]The 522 bp-length mitochondrial ND6 gene sequence of the S.curriculus,which encoded a 173 amino acid-length peptide,was obtained by PCR and sequencing in this research.It was found that the base composition of the ND6 gene was A 41.5%,C 32.7%,G 12.4 %,T 13.4%,respectively,and A + T content(54.9%) was higher than the G + C content(45.1%).Alignment of ND6 gene nucleotide sequence of S.curriculus with other 11 fish species from GenBank showed that the sequence homology was highest at 90.8% between S.curriculus and Megalobrama amblycephala,and lowest at 73.0% between S.curriculus and Salvelinus alpinus.Homology analyses on ND6 amino acid sequence showed that the homology was highest at 98.3% between S.curriculus and M.amblycephala,and lowest at 75.7% between S.curriculus and Larimichthys crocea.A phylogenetic tree constructed using the neighbor-joining method based on the ND6 amino acid sequence indicated that the relationship was in accord with morphologic classification.[Conclusion]The ND6 gene sequence of S.curriculus was successfully obtained.A phylogeny of S.curriculus and other 11 fish species was reconstructed.

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