青虾5个性别或生殖相关基因的克隆和表达分析
发布时间：2020-12-11 07:43
青虾,学名日本沼虾（Macrobrachium niponense） 广泛分布于全国各地。青虾是我国重要的淡水养殖虾类,养殖年产量约25万吨,养殖年产值超过100亿元。青虾雄性生长速度快于雌性,收获期规格也大于雌性。但由于迄今尚不了解青虾性别调控（涉及性别决定和性腺分化）的分子机制,建立大规模养殖全雄青虾的技术是困难的。正因为如此,开展青虾性别决定和性腺分化（生殖）的分子机制的研究十分重要。本实验已经构建青虾促雄性腺转录组和精巢、卵巢基因表达谱,为性别或生殖相关基因的筛选提供了理论指导。本文筛选了 Feminization-1 homolog b、Nitric oxide synthase、glycogen debranching enzyme、2 个 Serine protease inhibitor 共 5 个性别或生殖相关的候选基因,进行全长cDNA克隆及其时空表达模式分析,藉此确定其是否参与性别决定或生殖过程,以推动青虾性别调控分子机制的研究。Feminization-1 homolog b（Fem-1b）基因参与雄性动物的发育调控,在线虫性别控制中发挥着关键的作用,但在青虾中...
【文章来源】：南京农业大学江苏省 211工程院校 教育部直属院校
【文章页数】：128 页
【学位级别】：博士
【文章目录】：
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
【参考文献】：
期刊论文
[1]池蝶蚌致雄性化基因fem-1c及其蛋白分子的结构特征分析[J]. 熊文芳,史建伍,蒋谋炎,陈永玲,彭扣,盛军庆,王军花,洪一江. 水生生物学报. 2014(06)
[2]东亚飞蝗fem-1基因的克隆与表达分析[J]. 时红,郝友进,陈斌,司凤玲,王鹏,何正波. 昆虫学报. 2013(07)
[3]日本沼虾胚胎发育的形态及组织学观察[J]. 陈瑛,朱琴,陈辉,朱小玲,崔峥,邱高峰. 上海海洋大学学报. 2012(01)
[4]日本沼虾种质资源挖掘和保护研究进展[J]. 冯建彬,李家乐,程熙. 上海水产大学学报. 2008(03)
本文编号：2910147
【文章来源】：南京农业大学江苏省 211工程院校 教育部直属院校
【文章页数】：128 页
【学位级别】：博士
【文章目录】：
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
【参考文献】：
期刊论文
[1]池蝶蚌致雄性化基因fem-1c及其蛋白分子的结构特征分析[J]. 熊文芳,史建伍,蒋谋炎,陈永玲,彭扣,盛军庆,王军花,洪一江. 水生生物学报. 2014(06)
[2]东亚飞蝗fem-1基因的克隆与表达分析[J]. 时红,郝友进,陈斌,司凤玲,王鹏,何正波. 昆虫学报. 2013(07)
[3]日本沼虾胚胎发育的形态及组织学观察[J]. 陈瑛,朱琴,陈辉,朱小玲,崔峥,邱高峰. 上海海洋大学学报. 2012(01)
[4]日本沼虾种质资源挖掘和保护研究进展[J]. 冯建彬,李家乐,程熙. 上海水产大学学报. 2008(03)
本文编号：2910147
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