| ABSTRACT | 第14-19页 |
| 摘要 | 第20-24页 |
| LIST OF ABBREVIATIONS | 第24-26页 |
| PART ONE | 第26-64页 |
| CHAPTER ONE GENERAL INTRODUCTION AND LITERATURE REVIEW | 第26-55页 |
| 1.1 INTRODUCTION | 第26-28页 |
| 1.2 OBJECTIVES OF THIS STUDY | 第28-30页 |
| 1.2.1 General objectives | 第28页 |
| 1.2.2 Specific objective | 第28-30页 |
| 1.3 LITERATURE REVIEW | 第30-33页 |
| 1.3.1 Geese classification | 第30页 |
| 1.3.2 Chinese geese breeds | 第30页 |
| 1.3.3 Seasonality breeding of Chinese geese breeds | 第30-31页 |
| 1.3.4 Yangzhou Goose Breed | 第31页 |
| 1.3.5 Ovarian function and regulation in avian (geese) | 第31-32页 |
| 1.3.6 Granulosa cell | 第32-33页 |
| 1.4 ANIMAL TRAITS ASSESSMENT | 第33-34页 |
| 1.4.1 Linkage studies | 第33页 |
| 1.4.2 Candidate gene method | 第33-34页 |
| 1.4.3 Candidate gene implementation steps | 第34页 |
| 1.5 MELATONIN GENE | 第34-38页 |
| 1.5.1 Melatonin biosynthetic pathway | 第34-35页 |
| 1.5.2 Melatonin and Reproduction | 第35-36页 |
| 1.5.3 Melatonin as anti-oxidative in the follicle | 第36页 |
| 1.5.4 Melatonin Receptors | 第36-38页 |
| 1.5.4.1 Melatonin receptor subtype 1A | 第37-38页 |
| 1.6 EPIDERMAL GROWTH FACTOR | 第38-39页 |
| 1.7 EPIDERMAL GROWTH FACTOR RECEPTOR | 第39页 |
| 1.8 ENDOTHELIN GENE | 第39-40页 |
| 1.9 ENDOTHELIN RECEPTORS | 第40-41页 |
| 1.10 MICRORNA | 第41-42页 |
| 1.11 MIRNAS FUNCTIONS IN ANIMALS | 第42-43页 |
| REFERENCES | 第43-55页 |
| CHAPTER TWO APPLICATION OF PAIRED-END SEQUENCING TECHNOLOGY TO IDENTIFYSINGLE NUCLEOTIDE POLYMORPHISMS AND CANDIDATE GENES ASMARKERS ASSISTED SELECTION IN YANGZHOU GOOSE | 第55-64页 |
| 2.1 INTRODUCTION | 第55-56页 |
| 2.2 MATERIALS AND METHODS | 第56页 |
| 2.3 RESULTS | 第56-63页 |
| REFERENCE | 第63-64页 |
| PART TWO EXPERIMENT STUDY | 第64-185页 |
| CHAPTER ONE | 第64-102页 |
| SECTION ONE ASSOCIATION OF SINGLE NUCLEOTIDE POLYMORPHISM IN MELATONINRECEPTOR 1A GENE WITH EGG PRODUCTION AND MRNA EXPRESSION INYANGZHOU GEESE | 第64-85页 |
| 1.2 INTRODUCTION | 第65-66页 |
| 1.3 MATERIALS AND METHODS | 第66-71页 |
| 1.3.1 Experimental Birds, Blood Samples Collection and DNA Preparation | 第66-67页 |
| 1.3.2 PCR Amplification, Genotyping and SNP Validation | 第67-69页 |
| 1.3.3 Tissues Collection and Total RNA Isolation | 第69-70页 |
| 1.3.4 Quantitative real-time PCR Analysis of MTNR1A mRNA with SYBR GreenEnzyme | 第70页 |
| 1.3.5 Statistical Analysis | 第70-71页 |
| 1.4 RESULTS | 第71-78页 |
| 1.4.1 PCR products and sequence alignments | 第71-72页 |
| 1.4.2 Sequencing Analysis, Genotypes and Alleles Frequency Distribution | 第72-73页 |
| 1.4.3 Association Analysis of MTNR1A Polymorphisms with Total Number of EggProduction | 第73-74页 |
| 1.4.4 Distribution of mRNA Expression of MTNR1A Gene in Different Tissues andOvaries in two Groups (high and low) Egg Production from Yangzhou Geese | 第74-76页 |
| 1.4.5 Genetic Effects of (g.177G>C) SNP Polymorphism in MTNR1A mRNAExpression Levels in Ovary and Oviduct | 第76-78页 |
| 1.5 DISCUSSION | 第78-81页 |
| REFERENCES | 第81-85页 |
| SECTION TWO SINGLE NUCLEOTIDE POLYMORPHISMS IN PROMOTER REGION AFFECTTRANSCRIPTIONAL ACTIVITY OF THE MELATONIN RECEPTOR 1A GENEIN GEESE | 第85-102页 |
| 2.2 INTRODUCTION | 第85-87页 |
| 2.3 MATERIAL AND METHODS | 第87-92页 |
| 2.3.1 Experimental Materials | 第87页 |
| 2.3.2 Bioinformatics analysis | 第87页 |
| 2.3.3 Preparation of GG and CC polymorphism of Goose MTNR1A Gene Promoter | 第87-90页 |
| 2.3.4 Granulosa cell isolation | 第90-91页 |
| 2.3.5 Cell culture, transfections and Luciferase Activity Assay | 第91-92页 |
| 2.3.6 Statistical analysis | 第92页 |
| 2.4 RESULTS | 第92-97页 |
| 2.4.1 Bioinformatics analysis of MTNR 1A promoter region | 第92-93页 |
| 2.4.2 Luciferase Activity Analysis of MTNR 1 A gene SNP g.177G>C Fragments ofYangzhou goose | 第93-97页 |
| 2.4.2.1 Luciferase Activity Analysis of pGL3-314G, pGL3-314C and pGL3-basicvectors in HEK 293T cells | 第93-95页 |
| 2.4.2.2 Luciferase Activity Analysis of pGL3-314G, pGL3-314C and pGL3-basicvectors in geese granulosa cells | 第95-97页 |
| 2.5 DISCUSSION | 第97-99页 |
| REFERENCES | 第99-102页 |
| CHAPTER TWO | 第102-137页 |
| SECTION ONE A SNP AT MICRORNA BINDING SITE OF EPIDERMAL GROWTH FACTORRECEPTOR 3'-UTR ASSOCIATED WITH EGG PRODUCTION AND MRNAEXPRESSION IN YANGZHOU GEESE | 第102-122页 |
| 1.2 Introduction | 第103-104页 |
| 1.3 MATERIALS AND METHODS | 第104-108页 |
| 1.3.1 Ethics statement | 第104-108页 |
| 1.3.2 Experimental birds, blood samples collection and DNA extraction | 第104-105页 |
| 1.3.3 PCR Amplification,Sequencing and SNP identification | 第105-106页 |
| 1.3.4 Tissue collection, total RNA extraction and EGFR expression profile | 第106-108页 |
| 1.3.5 Statistical analysis | 第108页 |
| 1.4 RESULTS | 第108-116页 |
| 1.4.1 PCR amplification, sequencing analysis, Genotypes and Alleles frequencydistribution | 第108-111页 |
| 1.4.2 Association analysis of c.~*7750G>A SNP polymorphisms of EGFR gene withtotal egg production | 第111-113页 |
| 1.4.3 Distribution of mRNA expression of EGFR gene in various tissues and ovarianfollicles of Yangzhou geese | 第113页 |
| 1.4.4 Expression levels of EGFR gene in high and low egg production groups anddifferent genotypes | 第113-116页 |
| 1.5 DISCUSSION | 第116-119页 |
| REFERENCES | 第119-122页 |
| SECTION TWO SINGLE-NUCLEOTIDE POLYMORPHISMS C.~*7750G>A IN THE EPIDERMALGROWTH FACTOR RECEPTOR GENE 3'-UTR AFFECTS THE BINDING SITEAND PROMOTER ACTIVITY OF TARGET DME-MIR-5-3Q MIRNA INYANGZHOU GEESE | 第122-137页 |
| 2.2 INTRODUCTION | 第123-124页 |
| 2.3 MATERIALS AND METHODS | 第124-128页 |
| 2.3.1 Animals and Sample Collection | 第124页 |
| 2.3.2 Bioinformatics analysis of EGFR 3'-UTR | 第124-125页 |
| 2.3.3 Plasmid construction and miRNA mimics synthetics | 第125-127页 |
| 2.3.4 Geese granulosa cells extraction | 第127页 |
| 2.3.5 Activity analysis of EGFR 3'-UTR targeted by dme-miR-5-3q or hsa-miR-1248 | 第127-128页 |
| 2.3.6 Statistical analysis | 第128页 |
| 2.4 RESULTS | 第128-132页 |
| 2.4.1 MicroRNAs prediction | 第128-129页 |
| 2.4.2 Effects of c.~*7750G>A SNP genotypes of EGFR 3'-UTR on luciferase reporteractivity | 第129-132页 |
| 2.4.2.1 Effects on granulosa cells | 第129-130页 |
| 2.4.2.2 Effects on Hela cells | 第130-132页 |
| 2.5 DISCUSSION | 第132-134页 |
| REFERENCES | 第134-137页 |
| CHAPTER THREE THREE SINGLE NUCLEOTIDE POLYMORPHISMS (SNPS) IN SPERMADHESION MOLECULE 1 (SPAM1) GENE ASSOCIATED WITH EGGPRODUCTION IN YANGZHOU GOOSE | 第137-164页 |
| 1.2 INTRODUCTION | 第138-139页 |
| 1.3 MATERIAL AND METHODS | 第139-144页 |
| 1.3.1 Indigenous geese population,blood samples collection and DNA preparation. | 第139-140页 |
| 1.3.2 PCR Amplification, Genotyping and SNP Validation | 第140页 |
| 1.3.3 Sequence analysis, SNP detection, validation and genotyping | 第140-141页 |
| 1.3.4 RNA isolation, cDNA synthesis and quantitative real-time PCR Analysis ofSPAM1 mRNA | 第141-142页 |
| 1.3.5 Transcription factor binding sites Prediction in goose | 第142-143页 |
| 1.3.6 Promoter activity analysis using Luciferase activity assay | 第143-144页 |
| 1.3.6.1 Preparation and plasmid construction | 第143页 |
| 1.3.6.2 Geese granulosa cells extraction, culture, transfections and luciferase assay | 第143-144页 |
| 1.3.7 Statistical Analysis | 第144页 |
| 1.4 RESULTS | 第144-158页 |
| 1.4.1 PCR products, sequence analysis polymorphism identification, genotypes andalleles frequency distribution | 第144-148页 |
| 1.4.2 Genetic variation and linkage analysis of three SNPs located in 5'untranslatedregion and exon of the SPAM1 gene in Yangzhou geese | 第148-149页 |
| 1.4.3 Association analysis between g206 G>C,c123A>T and c159 A>G SNPspolymorphisms in SPAM1 gene with a total number of eggs production | 第149-152页 |
| 1.4.4 SPAM1 mRNA expression profile in Yangzhou goose tissues | 第152-153页 |
| 1.4.5 Expression levels of SPAM1 gene in oviducts and ovaries of high and low eggproduction groups from Yangzhou geese | 第153-154页 |
| 1.4.6 Expression levels of SPAM1 gene in oviducts and ovaries in different genotypesfrom Yangzhou geese | 第154-155页 |
| 1.4.7 Transcription Factor Binding Sites in the SPAM1 promoter region in goose | 第155页 |
| 1.4.8 Luciferase Activity Analysis results of SPAM1 gene of Yangzhou goose | 第155-158页 |
| 1.5 DISCUSSION | 第158-161页 |
| REFERENCES | 第161-164页 |
| CHAPTER FOUR ASSOCIATION OF SINGLE NUCLEOTIDE POLYMORPHISMS IN INTRONONE OF ENDOTHELIN RECEPTOR TYPE B WITH TOTAL EGGPRODUCTION TRAIT IN YANGZHOU GEESE | 第164-185页 |
| 1.2 INTRODUCTION | 第164-166页 |
| 1.3 MATERIAL AND METHODS | 第166-169页 |
| 1.3.1 Experimental geese and DNA extraction | 第166页 |
| 1.3.2 Primer designing and PCR amplification | 第166-167页 |
| 1.3.3 Sequence analysis and SNP detection | 第167-168页 |
| 1.3.4 Tissues collection, mRNA extraction and cDNA synthesis | 第168页 |
| 1.3.5 Quantitative real-time PCR Analyses | 第168-169页 |
| 1.3.6 Statistical analysis | 第169页 |
| 1.4 RESULTS | 第169-179页 |
| 1.4.1 Sequences variation analyses, genotyping and allele frequencies | 第169-174页 |
| 1.4.2 Genetic variation analysis of the 5 SNPs located in intron one of the EDNRBgene | 第174-175页 |
| 1.4.3 Association analysis of five SNPs polymorphisms of EDNRB gene with totalegg production | 第175-177页 |
| 1.4.4 Distribution of mRNA expression of EDNRB gene in various tissues | 第177-178页 |
| 1.4.5 Expression of EDNRB mRNA in two group of egg production from Yangzhougeese | 第178-179页 |
| 1.5 DISCUSSION | 第179-182页 |
| REFERENCES | 第182-185页 |
| GENERAL CONCLUSION | 第185-187页 |
| INNOVATION | 第187-188页 |
| ACKNOWLEDGEMENTS | 第188-189页 |
| LIST OF PUBLISHED,SUBMITTED AND UNDER PREPARATION PAPERS | 第189页 |
