【作者】 陈冬生；

【导师】 聂刘旺；

【作者基本信息】 安徽师范大学 ， 动物学， 2003， 硕士

【摘要】 本文由文献综述和研究报告两部分组成一、文献综述1、从Sox基因的发现、特点及功能三个方面概述Sox基因的研究现状。Sox基因含有一个能与DNA特异结合的高度保守HMG-box基序。在个体发育过程中，该基因广泛参与了动物的性别决定及多种组织器官的形成。2、从DMRT基因的发现及功能等方面综述DMRT基因的研究进展。DMRT基因是一类新发现的含有DM保守盒的转录调控因子。该基因参与调节动物的性别决定与分化等过程。3、综述了Sox基因与DMRT基因在性别决定中的相互作用关系。二、研究报告1、参照人SRY基因HMG-box保守区的序列，设计一对兼并引物，扩增了扬子鳄的Sox基因，并对扩增产物进行了克隆与测序。结果在雌雄中均筛选出8个不同的Sox基因，长度均为216bp，无性别特异性。其DNA序列和编码的氨基酸序列与人相应Sox基因的相似性分别为88%、88%、88%、89%、87%、93%、87%、93%和91%、91%、91%、96%、100%、98%、94%、96%。显示出进化上的高度保守性。2、参照人DMRT1基因DM盒保守区的序列，设计一对特异引物，扩增了扬子鳄的DMRT1基因，并对扩增产物进行了SSCP分析和测序。结果显示：(1)扬子鳄DMRT1基因的扩增片段与人DMRT1基因扩增片段的大小相同，均为140bp；(2)雌雄扬子鳄DMRT1基因扩增片段大小虽然与人相同，但其单链迁移率与人有较大差异，而雌雄个体间无差异。(3)测序结果表明，雌雄扬子鳄DMRT1基因的序列一致，且其DNA序列和编码的氨基酸序列与人相应DMRT1基因的相似性分别为87%和95%。显示出高度的保守性。本研究为探索扬子鳄的性别决定机制、Sox基因与DMRT基因的功能及其进化等提供了分子资料。更多还原

【Abstract】 This paper consists of two parts,which are as follows:PART ONE: reviews1. The discovery,characteristic and function of Sox genes were discussed. Sox genes encode a group of proteins that carry a DNA-binding HMG domain. In various phages of embryonic development, Sox genes play a key role in sex-determining and in organism formation.2. The findings and function of DMRT gene were reviewed. DMRT genes are a kind of transcription factor which possesses a common motif called the DM domain. DMRT genes are considered to be involved in sex determination and sex differentiation.3. The interaction between Sox genes and DMRT genes in sex determining and sex differentiation was discussed.PART TWO: studies1. After a pair of degenerate primers based on the conservative region, HMG-box,of Human SRY gene were used, eight different fragments were amplified from both female and male Alligator sinensis whose length were all 216bp, then they were cloned by pUCm-T Vector and sequenced.The result indicated that they showed high homology to Human SOX genes, the identities to Human SOX genes in the DNA sequence and the amino acid sequence were 88%、88%、88%、89%、87%、93%、87%、93% and 91%、91%、91%、96%、100%、98%、94%、96%, respectively, which may be concluded that Sox genes were highly conservative in phylogenesis.2. The DMRT1 gene of Alligator sinensis was amplified by using a pair of special primers which can amplify the conservative motif (DM domain) of human DMRT1 gene,and then it was sequenced. The length of amplification was 140bp in both male and female, which was consistent with that of human DMRT1 gene.The result of SSCP analysis showed that there were many differences in the DMRT1 gene sequence between Alligator sinensis and human,but no differences between male and female Alligator sinensis. The sequence analysis showed that the identities of Alligator sinensis DMRT1 gene to Human DMRT1 gene in the DNA sequence and the amino acid sequence were 87% and 95%, respectively. The results indicated that DMRT genes were highly conservative in phylogeny.Our work provides molecular data for the study of sex-determining mechanism, the function of Sox genes and DMRT genes ,and the conservation of Sox genes’ and DMRT genes’ evolution.更多还原