猪生殖细胞特异性基因Sohlh1及启动子的研究

【目的】探究猪生殖细胞特异性基因Sohlh1在幼年和成年各组织中的表达情况,分子遗传进化特点,及其启动子的核心区域和特异性调控区间。【方法】通过RT-PCR验证猪不同时期不同组织Sohlh1基因的表达情况,通过生物软件分析其分子遗传进化特点,构建启动子验证载体,通过双荧光素酶分析基因启动子的核心区域和特异性调控区间。【结果】Sohlh1基因在仔猪与成年猪睾丸和仔猪卵巢中都有表达,在成年猪卵巢不表达;通过多个物种进化分析,Sohlh1基因随着脊椎动物从低等到高等的演变也在进化;通过双荧光素酶分析构建的启动子验证载体,猪Sohlh1基因的启动子的核心区域在78 bp附近,特异性调控区间在771~2 981 bp之间。【结论】基于Sohlh1基因的进化分析,作为试验动物模型,猪可能更优于啮齿类动物,其组织特异性表达可能主要通过其启动子特异性调控区间调控。

Analysis of pig germ-cell special gene Sohlh1 and its promoter

Objective] To study the expression of germ-cell special gene Sohlh1 in different tissues of young and adult pigs, analyze the features of molecular genetic evolution of Sohlh1 gene, and determine the core region and specific regulatory region of the promoter.Method]The expressions of Sohlh1 gene in different pig tissues and different time were verified by RT-PCR.The features of molecular genetic evo-lution of Sohlh1 gene was analyzed using biology software, the promoter was constructed and confirmed by a verification vector, and the core region and specific regulatory region of the promoter were analyzed by dual luciferase assay.Result]Sohlh1 gene was expressed in testis of piglet and adult pig and ovary of piglet, and was not expressed in ovary of adult pig.Sohlh1 gene had evolved along with the evolution of lower to higher vertebrates based on phylogenetic analysis of SOHLH1 proteins in multiple species.The core region of the promoter of pig Sohlh1 gene was near 78 bp, and the specific regulatory region was be-tween 771 and 2 981 bp based on dual luciferase assay of the promoter verification vector.Conclusion]Pig could be a superior animal model compared with rodent based on Sohlh1 gene evolution analysis.The tissue specific expression of Sohlh1 gene may be mainly controlled by the specific regulatory region of its promoter.