论文说明
声明
CONTENTS
摘要
ABSTRACT
ABBREVIATIONS
CHAPTER 1 INTRODUCTION AND LITERATURES REVIEWS
1．1 INTRODUCTION AND HYPOTHESIS
1．1．1 Candidate genes approach for animal breeding and selection
1．1．2 The relationship of milk fat and milk SCS
1．1．3 MAP4K4 gene for inflammation and immunity study
1．1．4 DGAT1 gene for milk fat study
1．1．5 Hypothesis of proposed thesis
1．2 RESEARCH PROGRESSES OF DAIRY COW MASTITIS
1．2．1 Introduction of bovine mastitis and its impact on dairy industry
1．2．2 Genomic studies for bovine mastitis
1．3 Mechanism of pattern recognition receptors during mastitis
1．3．1 TLRs signaling in mastitis
1．3．2 Mechanism of RLRs during mastitis
1．3．3 NLRs and its expression during mastitis
1．4 AN INTRODUCTION OF MAP KINASE
1．4．1 MAP4K4 gene
1．4．2 MAPK pathways in the signaling network in mammalian cells
1．5 AN INTRODUCTION OF DGAT GENE
1．5．1 DGAT gene structure，ontogeny and expression pattern
1．5．2 DGAT1 gene and milk fat
1．6 OBJECTIVES OF THE PROPOSED RESEARCH AND INNOVATIONS
1．6．1．Experiment Ⅰ
CHAPTER 2 ASSOCIATION OF SINGLE NUCLEOTIDE POLYMORPHISMS IN MAP4K4 GENE WITH MASTITIS AND MILK TRAITS IN DAIRY COW
2．1 INTRODUCTION
2．2 MATERIALS AND METHODS
2．2．1 Ethics statement
2．2．2 Blood sample collection and genomic DNA extraction
2．2．3 Primer design and sequencing
2．2．4 Sequence analysis and genotyping
2．2．5 Statistical analysis
2．3 RESULTS
2．3．1 Characterization of Bovine MAP4k4(boMAP4K4)SNPs
2．3．2 Genotype and allelic frequencies
2．3．3 Analysis of boMAP4K4 SNPs with SCS
2．3．4 Association of boMAP4 SNPs with milk traits
2．4 DISCUSSION
CHAPTER-3 FUNCTION AND MECHANISMS OF DIFFERENT SEQUENCE OF MAP4K4 GENE BETWEEN DAIRY COW AND BUFFALO
3．2 MATERIALS AND METHODS
3．2．1 Animals samples and DNA extraction
3．2．2 Primer design，sequencing and SNP identification
3．2．3 Bioinformatics analysis and software used
3．2．4 Bovine mammary epithelium cell culture and LPS treatment
3．2．5 Extraction of RNA and cDNA，qPCR
3．2．6 Promoter cloning and generation of luciferase reporter constructs
3．2．7 Cell lines and culture conditions
3．2．8 Transient transfection and luciferase reporter assay
3．2．9 Statistical analysis
3．3 RESULTS
3．3．1．Structure characteristics of the bovine MAP4K4 gene
3．3．2 Difference screening in cow and buffalo sequence
3．3．3 Characterization of the bovine MAP4K4 gene 5'-regulatory region
3．3．4 Protein interaction analysis
3．3．5 Relative mRNA expression level analysis
3．3．6 Single nucleotide polymorphism and comparison
3．3．7 Construction of plasmid vectors
3．3．8 Isolation of the functional proximal minimal promoter of MAP4K4 gene
3．3．9 Comparison of functional minimal MAP4K4 promoter of cow and buffalo
3．4 DISCUSSION
CHAPTER-4 THE FUNCTIONS AND MECHANISMS OF SEQUENCE DIFFERENCES OF DGAT1 GENES ON MILK FAT IN BETWEEN DAIRY COW AND BUFFALO
4．2 MATERIALS AND METHODS
4．2．1 Animals samples and DNA extraction
4．2．2 Primer design，sequencing and SNP identification
4．2．3 Bioinformatics analysis and software used
4．2．4 Promoter cloning and generation of luciferase reporter constructs
4．2．5 Cell lines and culture conditions
4．2．6 Transient transfection and lueiferase reporter assay
4．2．7 Statistical Analysis
4．3 RESULTS
4．3．2 Difference screening in cow and buffalo DGAT1 sequence
4．3．3 Characterization of the bovine MAP4K4 gene 5'-regulatory region
4．3．4 Protein interaction analysis
4．3．5 Single nucleotide polymorphism and comparison
4．3．6 Construction of PLASMID VECTORS
4．3．7 Isolation of the functional proximal minimal promoter of DGAT1 gene
4．3．8 Comparison of the functional minimal DGAT1 promoter of cow and buffalo
4．4 DISCUSSION
CHAPTER 5 CONCLUSION AND FUTURE DIRECTION
REFERENCES
LIST OF FIGURE LEGENDS
LIST OF APPENDIX
ACKNOWLEDGEMENT
List of Publications
AUTHORS BIBLIOGRAPHY
