摘要
利用长片段PCR及T-A克隆技术,从兰州鲇(Silurus lanzhouensis)基因组DNA中首次克隆兰州鲇GH基因全序列并提交至Genbank获得登录号KM215221,同时,使用DNAstar等生物信息学软件进行序列分析研究。兰州鲇GH基因全长2 067 bp,由5个外显子和4个内含子组成,完整编码序列(Complete coding sequence,CDS)为603 bp。5个外显子大小分别为10、140、117、132和204 bp;4个内含子大小分别为229、103、565和103 bp;内含子与外显子的连接区序列遵循基因组成规则。编码区编码1条由200个氨基酸残基组成的蛋白质多肽。5个科8种鲶形目鱼类生长激素基因编码区序列同源性比较表明,它们之间GH基因编码区序列有较高的同源性,平均达到了92.4%,其中,兰州鲇和南方大口鲇之间的同源性最高,为99.5%,兰州鲇和革胡子鲇之间的同源性最低,为90.0%,并构建了系统发育进化树。同时也表明,GH基因编码区序列具有较高的保守性,系统发育研究中具有较高的参考价值。
Growth hormone（ GH） genes containing a complete open reading frame were cloned from genomic DNA of Silu-rus lanzhouensis using long PCR and T-A cloning techniques, and the accession numbers in GenBank is KM215221.At the same time, these sequence were analyzed by DNAstar and other bioinformatics software. The full-length of S.lanzhouensis GH gene was 2 067 bp, including five exons and four introns, and the complete coding sequence （ CDS） was 603 bp.The size of 5 exon size were 10 bp, 140 bp, 117 bp, 132 bp and 204 bp respectively and 4 intron were 229 bp, 103 bp, 565 bp and 103 bp respectively.The junction sequence of introns and exons of S.lanzhouensis GH gene was in accordance with the rules of gene composition.The coding region encoded a polypeptide with 200 amino acid residues. The homology comparison indicated that the average sequence homology reached 92.4% among 8 species of siluriformes catfish.Results of homology analysis revealed that the sequence of S.lanzhouensis GH gene shared the highest homology of 99.5%with the S.meridionals, while lowest 90.0%between S.lanzhouensis and Clarias lazera.Also the phylogenetic tree of 8 species were established according to sequence alignment results.The results also indicated that, the encoding of GH gene sequences were highly conservative and this study will provide a reference for further research.
出处
《淡水渔业》
CSCD
北大核心
2015年第4期20-25,共6页
Freshwater Fisheries
基金
国家自然科学基金项目(31360633)
宁夏对外科技合作项目
宁夏回族自治区自然科学基金(NZ14212)
