魚類性別控制研究是現(xiàn)代水產養(yǎng)殖的熱點。金錢魚（Scatophagus argus,Linnaeus,1766）是一種廣鹽性魚類,可作觀賞魚和食用魚,在養(yǎng)殖群體中性比為1:1。金錢魚雌魚的生長比雄魚快,個體大,有更好經濟價值。因此,克隆性別決定主效基因（master sex-determination gene,SD）將有助于金錢魚的性別控制。本研究旨在研究金錢魚的性別決定和分化機制,并為以后的單性（全雌魚）金錢魚培育提供理論基礎。下面為本研究的主要發(fā)現(xiàn):1、金錢魚性別特異性標記的開發(fā)和性別決定候選基因的克隆本研究克隆了金錢魚Dmrt1（Dmrt1和Dmrt1b）和Dmrt3（Dmrt3和Dmrt3△-Y）的兩個亞型。本研究開發(fā)了兩個位于金錢魚Dmrt3和Dmrt1上的性別連鎖分子標記,分別命名為Dmrt3-Marker-F/R和Dmrt1-Marker-F/R。Dmrt3-Marker-F/R擴增的片段包含能導致截短型Dmrt3（Dmrt3△-Y）c DNA提前編碼終止的基因組序列。Dmrt3-Marker-F/R擴增片段的151和166 bp單核苷酸多態(tài)性（SNP）顯示雌性個體均為C... 

【文章來源】：廣東海洋大學廣東省

【文章頁數】：121 頁

【學位級別】：碩士

【文章目錄】：
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


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期刊論文
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[2]金錢魚抗繆勒氏管激素基因克隆及其在性腺發(fā)育不同時期mRNA表達水平的分析[J]. 曾文剛,劉振浩,李紅,張俊彬.  水產學報. 2015(11)
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本文編號：3593443