利用两种标记技术分析中国海门鸡群的遗传多样性和遗传距离

摘要

本文利用RAPD和微卫星标记技术，评估了中国海门京海集团鸡群间的遗传多样性和遗传距离，同时也论证了这两种标记技术作为衡量群体杂合度(基因多样性)和其它群体基础遗传数据的准确工具的应用潜力，结果表明在中国江苏省海门京海集团4个鸡群(分别为RYC，JYC，WYC，CYC)中显示条带。从这4个鸡群共240个样本中(每个鸡群为60个样本)，每个群体随机抽取10个样本作为初始模板DNAs，并用上述60个10碱基大小的随机引物进行扩增，其中S246，S249，S1083，S1088和S1092这5条引物，占总引物的8.3333﹪，可以产生明显的多态性、可判断和可重复的条带，因此被作为标准引物而用于后续分析。　　本文认为，这两种遗传标记分析方法对于检测试验鸡群的遗传多样性和遗传距离都是有效可行的，然而，由于微卫星分析可以产生高度多态性，更高的杂合度并能检测到稀有基因，因此对于将来具有相对更大样本规模的鸡群遗传信息的研究微卫星标记分析方法是积极可行的。

目录

文摘

英文文摘

论文说明：DEDICATION、LIST OF TABLES、LIST OF FIGURES

CERTIFICATION

ACKNOWLEDGEMENTS

CHAPTER ONE

1.0 INTRODUCTION

CHAPTER TWO

2.0 LITERATURE REVIEW

2.1 GENETIC MARKERS IN THE CHICKENS

2.2 THE CHICKEN GENOME

2.3TECHNIQUES FOR THE MEASUREMENTS OF DIVERSITY IN ANIMAL AND POULTRY GENETIC RESOURCES

CHAPTER THREE

3.0 MATERIALS AND METHODS

3.1 EXPERIMENTAL STOCKS AND SAMPLE SIZE

3.2 BLOOD COLLECTION AND PROCESSING OF BLOOD SAMPLES

3.3 DNA ISOLATION AND PURIFICATION

3.4 DNA YIELD AND PURITY

3.5 RAPD-PCR PROTOCOLS AND AGAROSE GEL ELECTROPHORESIS

3.5.1 PCR conditions and programme

3.5.2 Submerged agarose gel electrophoresis (SAGE)

3.6 PREPARATION OF POOLED DNA SAMPLES AND RAPD-PCR PROTOCOL OF THE BULKED SAMPLES

3.7 MICROSATELLITE ANALYSIS

3.8 MICROSATELLITE PCR PROTOCOL AND ELECTROPHORESIS

3.8.1 Polyacrylamide gel electrophoresis (PAGE)

3.9 STATISTICAL ANALYSES

CHAPTER FOUR

4.0 RESULTS

4.1 PROPORTION OF POLYMORPHISMS IN POPULATIONS

4.2 AMPLIFICATION RESULTS OF POPULATION POOLED DNAs

4.3 BAND SHARING COEFFICIENTS AND GENE DIVERSITY VALUES OF THE PAIRWISE CHICKEN POPULATIONS

4.4 AVERAGE PAIRWISE PERCENT DIFFERENCE (APD) AMONG THE FOUR CHICKEN POPULATIONS

4.5 GENETIC DISTANCES AMONG THE FOUR CHICKEN POPULATIONS USING RAPD MARKERS

4.6 MICROSATELLITE LOCI ANALYSIS AND RESULTS

4.6.1 Rare/uncommon/private alleles

4.7 GENETIC DIVERSITY (VARIABILITY) AMONG THE CHICKEN POPULATIONS EXAMINED

4.7.1 Allele frequency

4.7.2 Heterozygosity

4.7.3 Effective number of allele and polymorphism information content

4.8 GENETIC DIVERGENCE/DIFFERENTIATION AND SHANNON INFORMATION INDEX

4.9 GENETIC DISTANCES

CHAPTER FIVE

5.0 DISCUSSION

5.1 RAPD DNA FRAGMENT BANDS

5.2 BAND SHARING COEFFICIENT, HETEROZYGOSITY AND GENETIC DISTANCE

5.3 MICROSATELLITES ALLELE FREQUENCY INTERPRETATION

5.4 HETEROZYGOSITY, EFFECTIVE ALLELE NUMBER AND POLYMORPHISM INFORMATION CONTENT

5.5 PHYLOGENETIC RELATIONSHIPS

5.6 COMPARISONS OF THE TWO ASSAY METHODS USED IN THIS STUDY

5.7 FUTURE LINE OF STUDY

CHAPTER SIX

6.0 SUMMARY, CONCLUSION AND RECOMMENDATIONS

REFERENCES

PAPERS IN PEER-REVIEWED JOURNALS WITHIN THE STUDY PERIOD

RECENTLY SUBMITTED PAPERS FOR PUBLICATION

ADDENDUM