青蝦,學(xué)名日本沼蝦（Macrobrachium niponense） 廣泛分布于全國各地。青蝦是我國重要的淡水養(yǎng)殖蝦類,養(yǎng)殖年產(chǎn)量約25萬噸,養(yǎng)殖年產(chǎn)值超過100億元。青蝦雄性生長速度快于雌性,收獲期規(guī)格也大于雌性。但由于迄今尚不了解青蝦性別調(diào)控（涉及性別決定和性腺分化）的分子機(jī)制,建立大規(guī)模養(yǎng)殖全雄青蝦的技術(shù)是困難的。正因?yàn)槿绱?開展青蝦性別決定和性腺分化（生殖）的分子機(jī)制的研究十分重要。本實(shí)驗(yàn)已經(jīng)構(gòu)建青蝦促雄性腺轉(zhuǎn)錄組和精巢、卵巢基因表達(dá)譜,為性別或生殖相關(guān)基因的篩選提供了理論指導(dǎo)。本文篩選了 Feminization-1 homolog b、Nitric oxide synthase、glycogen debranching enzyme、2 個 Serine protease inhibitor 共 5 個性別或生殖相關(guān)的候選基因,進(jìn)行全長cDNA克隆及其時空表達(dá)模式分析,藉此確定其是否參與性別決定或生殖過程,以推動青蝦性別調(diào)控分子機(jī)制的研究。Feminization-1 homolog b（Fem-1b）基因參與雄性動物的發(fā)育調(diào)控,在線蟲性別控制中發(fā)揮著關(guān)鍵的作用,但在青蝦中... 

【文章來源】：南京農(nóng)業(yè)大學(xué)江蘇省 211工程院校 教育部直屬院校

【文章頁數(shù)】：128 頁

【學(xué)位級別】：博士

【文章目錄】：
ABSTRACT
摘要
List of abbreviation
CHAPTER 1 Literature review and study plan
    1. Definition of aquaculture
    2. Crustaceans
    3. Macrobrachium nipponense distribution and aquaculture
    4. Overview of aquatic animals Genetics
    5. Sex management and breeding
    6. Future evolution
    7. Needs for Sex Control
    8. Sex determination and sex differentiation in fish
    9. Approaches of Sex Manipulation
    10. Gene research in crustaceans
    11. Genetics and genes research in Macrobrachium nipponense
    12. Androgenic gland
    13. Justification of the study
    14. Aim of the study
    15. Hypothesis of the study
    16. Content of the study, gene selection and study schematic design
CHAPTER 2 Molecular cloning and expression analysis of Fem1b from Oriental River prawnMacrobrachium nipponense
    1 Introduction
    2 Materials and methods
        2.1 Prawn and tissue preparation
        2.2 RNA isolation and reverse transcription
        2.3 Rapid amplification of cDNA ends （RACE）
        2.4 Nucleotide sequence and bioinformatics analyses
        2.5 Real-time quantitative PCR analysis of Femlb
        2.6 Statistical analysis
    3 Results
        3.1 Sequence analysis of MnFem1b
        3.2 Homology and phylogenetic analysis of MnFem1b
        3.3 Tissue distribution of MnFem1b mRNA
        3.4 Expression analysis of MnFem1b mRNA during embryo,larvae,andpost-larval stages
    4 Discussion
    5 Conclusion
CHAPTER 3 Molecular cloning and expression patter of Nitric Oxide Synthase （NOS） in oriental riverprawn, Macrobrachium nipponense
    1 Introduction
    2 Materials and Methods
        2.1 Experimental Prawn and tissue collection
        2.2 RNA extraction and cDNA synthesis
        2.3 5'-and 3'-random amplification of cDNA ends （RACE） of NOS gene
        2.4 Analyses of the nucleotide and deduced amino acid sequence
        2.5 Expression of MnNOS in different adult tissues and developmental stages
        2.6 Statistical analysis
    3 Results
        3.1 Characterization of the full-length of MnNOS
        3.2 Phylogenetic analysis of MnNOS
        3.3 Expression of MnNOS mRNA in tissues
        3.4 Temporal expression of MnNOS mRNA during the different developmentalstages of prawn larvae, embryo and post larvae
    4 Discussion
    5 Conclusion
CHAPTER 4 Molecular Cloning and Expression Profile of Glycogen Debranching Enzyme-like Geneduring Developmental Stages of Macrobrachium nipponense（de Haan 1849）
    1 Introduction
    2 Material and methods
        2.1 Prawn and tissue preparation
        2.2 RNA isolation and reverse transcription
        2.3 Rapid amplification of cDNA ends of MnAGL （RACE）
        2.4 Nucleotide sequence and bioinformatics analyses
        2.5 Real-time quantitative PCR analysis of MnAGL
        2.6 Statistical analysis
    3 Result
        3.1 Structural analysis of a full-length cDNA of MnAGL
        3.2 Homology and phylogenetic analysis of AGL
        3.3 Phylogenetic analysis of MnAGL
        3.4 Tissue distribution of the MnAGL gene transcript
        3.5 Temporal expression of MnAGL mRNA during the early embryo,larvae andpost-larvae stages
    4 Discussion
    5 Conclusion
CHAPTER 5 Molecular cloning of two Serine protease inhibitor genes and expression analysis indifferent tissues in oriental river prawn,Macrobrachium nipponense
    1 Introduction
    2 Materials and methods
        2.1 Shrimp
        2.2 Total RNA extraction and first-strand cDNA synthesis
        2.3 Rapid amplification of cDNA ends （RACE）
        2.4 Tissue distribution analysis
        2.5 Nuleotide sequence and Bioinformatics analysis
    3 Results
        3.1 Sequence analysis of MnSERPIN1 and MnSERPIN2
        3.2 Bioinformatics analysis
        3.3 Homologous and phylogenetic analysis of MnSERPIN1 and MnSERPIN2
        3.4 Tissue expression of MnSERPIN1 and MnSERPIN2
    4 Discussion
    5 Conclusion
Conclusion of the whole study
References
Acknowledgements
Publications
Academic award
Academic seminars attended


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期刊論文
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本文編號：2910147