猪生殖细胞特异性基因Sohlh1及启动子的研究  

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

作　　者：刘凯[1] 刘国乾[1,2] 曹亚鸽 卫恒习[1] 李莉 张守全[1] 

机构地区：[1]华南农业大学动物科学学院/国家生猪种业工程技术研究中心/广东省农业动物基因组学与分子育种重点实验室,广东广州510642 [2]广东科贸职业学院,广东广州510640

出　　处：《华南农业大学学报》2016年第6期52-58,共7页Journal of South China Agricultural University

基　　金：973计划项目(2011CB944202);国家自然科学基金(31572397)

摘　　要：【目的】探究猪生殖细胞特异性基因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】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78bp附近,特异性调控区间在771-2981bp之间.【结论】基于基因的进化分析,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.【 Objective】 To study the expression of germ-cell special gene Sohlh1 in different tissues ofyoung and adult pigs, analyze the features of molecular genetic evolution of Sohlh 1 gene, and determinethe core region and specific regulatory region of the promoter.【 Method 】 The expressions of SO/ hl genein different pig tissues and different time were verified by RT- PCR. The features olution of Sohlh 1 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 ofpiglet, and was not expressed in ovary of adult pig. Sohlhl gene had evolved along with the evolution oflower to higher vertebrates based on phylogenetic analysis of SOHLH 1 proteins in multiple species. The core region of the promoter of pig Sohlh 1 gene was near 7 8 bp, and the specific regulatory region was between 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 Sohlh 1 gene etissue specific expression of Sohlhl gene may be mainly controlled by the specific regulatory region of itspromoter.

关 键 词：猪 Sohlh1 启动子 遗传进化 验证载体 双荧光素酶 

分 类 号：S828[农业科学—畜牧学]