论文部分内容阅读
采用3’-和5’-RACE法克隆了泥鳅和大鳞副泥鳅CYP17-Ⅰ基因的c DNA全长序列,通过实时荧光定量PCR技术分析其表达情况。结果表明,泥鳅CYP17-Ⅰ基因c DNA全长1 706 bp,开放阅读框(open reading frame,ORF)1 563 bp,编码520个氨基酸;大鳞副泥鳅CYP17-Ⅰ基因c DNA全长1 763 bp,ORF长1 545 bp,编码514个氨基酸。2种鳅CYP17-Ⅰ氨基酸序列都有1个信号肽、1个跨膜区、1个保守的蛋白结构域和3个功能保守区。相似度分析显示,2种鳅之间CYP17-Ⅰ相似度为99%,与其他鱼类的相似度也超过70%。系统进化分析显示,2种鳅之间关系最为接近,其系统发育关系基本符合传统的分类地位。qRT-PCR结果显示,CYP17-Ⅰ在2种鳅的肠、肌肉、心脏、胃、肝脏、精巢、卵巢、脾脏等8个组织均有表达,在精巢和卵巢中表达量相对较高。
The full-length cDNA sequence of CYP17-Ⅰ gene from loach and Paramisgurnus dabryanus was cloned by 3’- and 5’-RACE, and its expression was analyzed by real-time fluorescence quantitative PCR. The results showed that the c DNA of CYP17-Ⅰ gene of loach was 1 706 bp in length and 1 563 bp in open reading frame (ORF), which encoded 520 amino acids. The ORF is 1 545 bp in length and encodes 514 amino acids. Two kinds of loach CYP17-Ⅰ amino acid sequence has a signal peptide, a transmembrane region, a conserved protein domain and three functionally conserved regions. The similarity analysis showed that the similarity of CYP17-Ⅰ between two species of loach was 99%, and the similarity with other fish species was more than 70%. Phylogenetic analysis showed that the relationship between the two species of Loach was the closest, and its phylogenetic relationship basically conformed to the traditional taxonomic status. The results of qRT-PCR showed that CYP17-Ⅰ was expressed in 8 tissues of intestine, muscle, heart, stomach, liver, testis, ovary and spleen of 2 species of loach and higher in both testis and ovary.