金钱鱼DMRT1和GSDF在性别决定与分化中的功能初步研究

发布时间：2022-01-16 21:21

　　鱼类性别控制研究是现代水产养殖的热点。金钱鱼（Scatophagus argus,Linnaeus,1766）是一种广盐性鱼类,可作观赏鱼和食用鱼,在养殖群体中性比为1:1。金钱鱼雌鱼的生长比雄鱼快,个体大,有更好经济价值。因此,克隆性别决定主效基因（master sex-determination gene,SD）将有助于金钱鱼的性别控制。本研究旨在研究金钱鱼的性别决定和分化机制,并为以后的单性（全雌鱼）金钱鱼培育提供理论基础。下面为本研究的主要发现:1、金钱鱼性别特异性标记的开发和性别决定候选基因的克隆本研究克隆了金钱鱼Dmrt1（Dmrt1和Dmrt1b）和Dmrt3（Dmrt3和Dmrt3△-Y）的两个亚型。本研究开发了两个位于金钱鱼Dmrt3和Dmrt1上的性别连锁分子标记,分别命名为Dmrt3-Marker-F/R和Dmrt1-Marker-F/R。Dmrt3-Marker-F/R扩增的片段包含能导致截短型Dmrt3（Dmrt3△-Y）c DNA提前编码终止的基因组序列。Dmrt3-Marker-F/R扩增片段的151和166 bp单核苷酸多态性（SNP）显示雌性个体均为C... 

【文章来源】：广东海洋大学广东省

【文章页数】：121 页

【学位级别】：硕士

【文章目录】：
Abbreviations
Abstract
摘要
1 General Introduction
    1.1 Background of the Study
    1.2 Literature Review on Teleost Sex Determination and Differentiation (SDD)
        1.2.1 Processes of Gonad Development in Gonochoristic
        1.2.2 Sex-Determination (SD) Genes in Fish
        1.2.3 Other Sex Related Genes
        1.2.4 Epigenetic Mechanisms
        1.2.5 Environmental Effects (EE) on Sex Determination and Differentiation (SDD)
        1.2.6 Understanding the Sex-Determination (SD) Pathway
        1.2.7 The Tag of War Between GSD and ESD
        1.2.8 Sex Control in Fish
    1.3 Problem Statement
    1.4 The Aim and Scope of the Study
    1.5 The Scientific Significance of the Study
2 Identification of Sex-Specific Markers and Cloning of Candidate Sex-Determination Gene in Spotted scat, Scatophagus argus
    2.1 Introduction
    2.2 Materials and Methods
        2.2.1. Animals and Samples Collection
        2.2.2 RNA Extraction
        2.2.3 Genomic DNA (gDNA) Extraction
        2.2.4 Cloning of cDNA and gDNA
        2.2.5 Sequence Analysis
        2.2.6 Syntenic Analysis
        2.2.7 Reverse Transcription PCR (RT-PCR)
    2.3 Results
        2.3.1 cDNA Cloning and Sequence Analysis
        2.3.2 Phylogenetic Analysis
        2.3.3 Screening of Sex-Specific Marker on Dmrt3 Locus
        2.3.4 Cloning of Male-Specific Dmrt1
        2.3.5 Synthenic Analysis
        2.3.6 Gene Expression Profile of Dmrt1, Dmrt1b, Dmrt3 and Dmrt3△-Y in Gonads of Scatophagus argus
    2.4 Discussion
    2.5 Conclusion
3 Expression Analysis and Relationship Study of Dmrt1 and Gsdf in Spotted Scat(Scatophagus argus)
    3.1 Introduction
    3.2 Materials and Methods
        3.2.0 Animals and Sampling
        3.2.1 Genotyping and Phenotyping of the Sex
        3.2.2 H&E Staining
        3.2.3 cDNA Cloning and Sequence Analysis
        3.2.4 Tissue Distribution of gsdf, dmrt1 and sf1 mRNA
        3.2.5 Expression of gsdf, dmrt1 and sf1 at Different Gonad Developmental Phases
        3.2.6 Western Blotting
        3.2.7 Immunohistochemistry (IHC)
        3.2.8 Primary Antibody Pre-adsorption Test
        3.2.9 In Silico and In Vitro Promoter Analysis
        3.2.10 Statistical Analysis
    3.3 Results
        3.3.1 Fish Sampling
        3.3.2 cDNA Cloning and Sequence Analysis
        3.3.3 Tissue Distribution
        3.3.4 Expression of gsdf, dmrt1 and sf1 mRNA at Different Gonad Developmental Phases
        3.3.5 Gsdf Protein Expression in Gonads by Western Blot
        3.3.6 Expression of Gsdf at Different Gonad Developmental Phases by IHC
        3.3.7 Transcriptional Regulation of gsdf
    3.4 Discussion
        3.4.1 Gsdf Showed Sex-biased Expression in Fish and Activated by Dmrt1 in Male Spotted Scat
        3.4.2 Function of Gsdf in Ovary Development and Other Possible Factors Influence Gsdf Expression
    3.5 Conclusions
4 General Conclusion and Recommendations
    4.1 Conclusion
    4.2 Recommendations
References
Acknowledgements
Author’s profile
Supervisors’ profile


【参考文献】：
期刊论文
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[2]金钱鱼抗缪勒氏管激素基因克隆及其在性腺发育不同时期mRNA表达水平的分析[J]. 曾文刚,刘振浩,李红,张俊彬.  水产学报. 2015(11)
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