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Sox9基因是一个重要的转录调控因子,参与性别决定及软骨等多种组织和器官的发育过程。本研究利用简并引物扩增鲤鱼基因组DNA,首次发现在鲤鱼中存在两种形式的Sox9基因。二者在保守盒区编码的氨基酸序列相同,并都存在一个内含子,但内含子序列差异很大,分别长704bp和616bp。在此基础上采用RACE技术克隆了鲤鱼Sox9b基因的5’端和3’端,通过拼接获得了2447bp 的全长cDNA序列,编码428个氨基酸。其中96-174位共79个氨基酸为HMG保守盒。将鲤鱼Sox9b基因与三刺鱼等九种动物的氨基酸序列相比较发现,它们的同源性高达75%以上,显示Sox9 基因在进化中较保守。应用半定量RT-PCR技术对成体鲤鱼不同组织中Sox9b基因的表达进行了分析,结果表明该基因广泛表达,尤以脑及精巢中表达最为丰富。
Sox9 gene is an important transcriptional regulator involved in sex determination and the development of various tissues and organs such as cartilage. In this study, degenerate primers were used to amplify common carp genome DNA. For the first time, two forms of Sox9 gene were found in common carp. The two encoded the same amino acid sequence in the conserved box region, and both contained an intron. However, the intron sequences varied greatly, by 704 bp and 616 bp, respectively. On this basis, the 5’-end and the 3’-end of Sox9b gene were cloned by RACE technology. The full-length cDNA sequence of 2447 bp was obtained by splicing, encoding 428 amino acids. A total of 79 amino acids from 96 to 174 were HMG conserved boxes. Comparing the amino acid sequences of carp Sox9b gene with nine kinds of animals, such as tuna, found that their homology was as high as 75%, indicating that Sox9 gene is more conservative in evolution. The expression of Sox9b gene in different tissues of adult carp was analyzed by semi-quantitative RT-PCR. The results showed that the gene was widely expressed, especially in brain and testis.