青蝦5個性別或生殖相關基因的克隆和表達分析
發(fā)布時間：2020-12-11 07:43
青蝦,學名日本沼蝦（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控（涉及性別決定和性腺分化）的分子機制,建立大規(guī)模養(yǎng)殖全雄青蝦的技術是困難的。正因為如此,開展青蝦性別決定和性腺分化（生殖）的分子機制的研究十分重要。本實驗已經(jīng)構建青蝦促雄性腺轉錄組和精巢、卵巢基因表達譜,為性別或生殖相關基因的篩選提供了理論指導。本文篩選了 Feminization-1 homolog b、Nitric oxide synthase、glycogen debranching enzyme、2 個 Serine protease inhibitor 共 5 個性別或生殖相關的候選基因,進行全長cDNA克隆及其時空表達模式分析,藉此確定其是否參與性別決定或生殖過程,以推動青蝦性別調控分子機制的研究。Feminization-1 homolog b（Fem-1b）基因參與雄性動物的發(fā)育調控,在線蟲性別控制中發(fā)揮著關鍵的作用,但在青蝦中...
【文章來源】：南京農業(yè)大學江蘇省 211工程院校 教育部直屬院校
【文章頁數(shù)】：128 頁
【學位級別】：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
【參考文獻】：
期刊論文
[1]池蝶蚌致雄性化基因fem-1c及其蛋白分子的結構特征分析[J]. 熊文芳,史建伍,蔣謀炎,陳永玲,彭扣,盛軍慶,王軍花,洪一江. 水生生物學報. 2014(06)
[2]東亞飛蝗fem-1基因的克隆與表達分析[J]. 時紅,郝友進,陳斌,司鳳玲,王鵬,何正波. 昆蟲學報. 2013(07)
[3]日本沼蝦胚胎發(fā)育的形態(tài)及組織學觀察[J]. 陳瑛,朱琴,陳輝,朱小玲,崔崢,邱高峰. 上海海洋大學學報. 2012(01)
[4]日本沼蝦種質資源挖掘和保護研究進展[J]. 馮建彬,李家樂,程熙. 上海水產(chǎn)大學學報. 2008(03)
本文編號：2910147
【文章來源】：南京農業(yè)大學江蘇省 211工程院校 教育部直屬院校
【文章頁數(shù)】：128 頁
【學位級別】：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
【參考文獻】：
期刊論文
[1]池蝶蚌致雄性化基因fem-1c及其蛋白分子的結構特征分析[J]. 熊文芳,史建伍,蔣謀炎,陳永玲,彭扣,盛軍慶,王軍花,洪一江. 水生生物學報. 2014(06)
[2]東亞飛蝗fem-1基因的克隆與表達分析[J]. 時紅,郝友進,陳斌,司鳳玲,王鵬,何正波. 昆蟲學報. 2013(07)
[3]日本沼蝦胚胎發(fā)育的形態(tài)及組織學觀察[J]. 陳瑛,朱琴,陳輝,朱小玲,崔崢,邱高峰. 上海海洋大學學報. 2012(01)
[4]日本沼蝦種質資源挖掘和保護研究進展[J]. 馮建彬,李家樂,程熙. 上海水產(chǎn)大學學報. 2008(03)
本文編號：2910147
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