西伯利亚鲟Sox17基因cDNA全长的克隆及其表达分析

Cloning,analysis and expression detection of full-length c DNA of Sox17 from Acipenser baerii

采用RACE技术获得西伯利亚鲟(Acipenser baerii)Sox17基因cDNA全长,序列分析表明,西伯利亚鲟Sox17基因cDNA全长2 671 bp,包括588 bp的5'端非翻译区,901 bp的3'端非翻译区和编码393个氨基酸残基的1 183 bp开放阅读框。氨基酸序列比对显示,该基因具较高的保守性,与塞内加尔多鳍鱼(Polypterus senegalus)的相似度最高为72%。系统进化分析表明,西伯利亚鲟与塞内加尔多鳍鱼聚为一支,且支持率为95%。以延伸因子efla(elongation factor 1-alpha)作为内参基因,对Sox17基因在西伯利亚鲟各组织的相对表达量进行分析,发现Sox17基因在不同组织中均有表达,但表达具有明显的组织差异性。在脑中表达量最高,而在肾、血液、眼、肠和胸鳍5个组织中表达量均较低。同时,Sox17基因在西伯利亚鲟不同发育时期均有表达,但表达具有一定的时间差异性。在宽神经板期的表达量高,相当于表达量最低点11日龄仔鱼的167倍。本研究为量化西伯利亚鲟胚胎发育分化过程中相关基因提供了参照工具,同时可为今后深入研究西伯利亚鲟Sox17在胚胎发育过程中的作用提供基础参考资料。

In this study, the full-length cDNA of Soxl7 gene from Siberian sturgeon （Acipense baerii） was obtained using RACE techniques. The full-length of Soxl 7 gene cDNA was 2 671 bp in length,including a 1 183 bp open reading frame（ORF） encoded 393 amino aeids,a 588 bp 5 untranslated region （5＇ UTR） and a 901 bp 3 untranslated region（3＇ UTR）. Multiple alignment analyses showed Siberian sturgeon was highly conservative to other fishes. It had high similarity with Ploypterus senegalus of 72% support rating. Phylogenetic analysis showd that Siberian sturgeon first clustered together with Ploypterus senegalus with the approval rating of 95 %. And then efla （ elongation factor 1-alpha, efla） gene was used as a internal reference on detecting the expression of Soxl7 gene in various tissues of Siberian sturgeon by real-time fluorescence quantitative PCR. The results showed Soxl7 gene expressed in diferent tissues and the expression level of Soxl7 was relatively low in kidney, blood, eye, intestine and pelvic fin. However, in brain, the expression level of Soxl 7 was the highest in all of these tissues. At the same time, Soxl 7 gene also expressed in different development periods, but expressed with a certain amount of time difference. In wide neural plate, the expression level of Soxl7 was the highest, reached 167 times that of the quantity of 11 days old larvae period. This study provides a reference to quantify the Siberian sturgeon differentiation genes involved in the process of embryonic development. At the same time, it provides the basic reference for the future study of the Siberian sturgeon Soxl 7 role in the process of embryonic development.