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绵羊多产性状的DNA分子标记及相关研究

作　者: 管峰
导　师: 杨利国；刘守仁
学　校: 南京农业大学
专　业: 动物遗传育种与繁殖
关键词: Sheep BMPR-IB gene BMP 15 gene GDF9 gene Microsatellite markers Tetra-primer ARMS PCR Detection litter size
分类号: S826.2
类　型: 博士论文
年　份: 2005年
下　载: 305次
引　用: 4次
阅　读: 论文下载

内容摘要

Techniques such as candidate genes, microsatellite marker linkage analysis were used to study the frequency of prolific genes and the relationship among litter size, growth and development in seven sheep breeds or strains. A rapid and simple method for detection of FecB gene was established in this study. The purpose of the current study is to provide a method for molecular marker assistant breeding. The main contents presented as follows:1. Study of candidate genes on sheep prolificacyDNA samples were collected and amplified for bone morphogenetic protein receptor type IB gene (BMPR-IB), BMP 15 gene and GDF genes by PCR-RFLP method from 553 sheep from 7 breeds or strains such as Hu, Chinese Merino meat-Prolificacy strain, Suffolk, Dorset, Charolais, Chinese Merino and Romney Hills. Furthermore, relationship between the polymorphism of BMPR-IB gene and prolificacy as well as growth and development were analyzed. The results showed that the genotype frequency of BMPR-IB gene was significantly different in these breeds or strains. In Chinese Merino meat-prolificacy strain, the genotype frequency of BB, B+ and ++ is 51% 30% and 19% respectively. Except for Hu sheep with only BB genotype, other breeds had only ++ genotype. Analyzing the litter size, body weight and body size within Chinese Merino meat-Prolificacy strain, we found show that the mean litter size at 1st lambing of BB, B+ and ++ genotype populations is 2.0, 1.5 and 1.1 respectively, and there are significantly difference between groups (BB vs B+, P<0.05; BB vs ++, P<0.01). Sheep with genotype BB and B+ had a greater total litter size than those with genotype ++ (P<0.01), the number is 2.8, 2.3 and 1.2 respectively. At ages of 90 days, the body weight of lambs with genotype BB and B+ were (18.6±3.70)kg and ( 18.0 ± 3.31 ) kg, which were significantly greater than that of population with ++ (15.6 + 2.22 kg) genotype (p<0.05). Furthermore, at this age, the lambs with genotype BB and B+ were significantly greater than those with genotype ++ in chest circumstance and chest width, but at age of 120 days there was no significant difference between these two groups.These findings indicate that the BMPR-IB gene is a major gene that regulates litter size and affects growth and development during a certain period.2 . Study on polymorphisms of microsatellites in different sheeppopulationsSix microsatellite markers named LscvO43, BMS2508, GC101, 300U, Bulge5 and 471U which were closely linked to the FecB gene were analyzed in samples of Hu collected from Natural Source Conservative Region and Shanghai.for genetic diversity and relationship on litter size in Hu sheep as candidate markers. And Chinese Merino prolific meat strain as control. To evaluate the Hu breed by comparison with polymorphism information content (PIC), genetic heterozygosity (h), index of genetic diversity (H), and effective number of allele (E). The results showed that six markers were all polymorphic markers within Hu sheep, and total mean PIC, H, h and E within Hu come from Natural Source Conservative Region were higher than that of from Shanghai, but was lower than that of Chinese Merino prolific meat strain. The genetic distance (D) and index of similarity (I) of three different sheep population is in according with its breeding process. The mean litter size at 1st lambing for LscvO43 site HObp/llObp was the highest and total mean litter size for 300U site 148bp/148bp was the highest within Hu sheep population. Total mean litter size for LscvO43 site 107bp/123bp was significantly higher than that for LscvO43 site H0bp/123bp (PO.05). The 1st mean litter size for BMS2508 site 154bp/154bp, 154bp/170bp and 154bp/200bp was significantly higher than that for BMS2508 site 170bp/170bp (PO.05). There was difference between the mean litter size at 1st lambing and total mean litter size of microsatellite markers GC101, 300U, 471U and Bulge5 but no significantly difference was detected.3. Development of a method using tetra-primer ARMS PCR for detection of FecB gene mutation in sheepTetra-primer ARMS PCR is a rapid, simple and efficient method to detect single nucleotide polymorphism (SNP). Bone morphogenetic protein receptor IB (BMPR-IB) gene was a major effect gene that control prolific trait in Booroola Merino sheep. To establish a tetra-primer ARMS PCR method to genotype BMPR-IB gene. Specific primers around the mutation locus (A746G) were designed on the basis of tetra-primer

全文目录

英文摘要  7-10
引言  10-12
第一部分文献综述  12-43
  绵羊多产性分子标记研究进展  12-43
    1 遗传标记简介  12-24
      1.1 RFLP标记  12-13
      1.2 微卫星DNA  13-22
      1.3 单核苷酸多态性  22-24
    2 与绵羊繁殖性状有关的候选基因  24-36
      2.1 FecB基因  25-27
      2.2 FecX基因  27-28
      2.3 BMP15和GDF9基因  28-30
      2.4 其它多胎基因  30-32
      2.5 各种突变的相互作用  32-33
      2.6 BMP与绵羊多胎机理  33-36
    3 Tetra-ARMS PCR技术研究进展  36-39
      3.1 错配PCR基本原理及衍生SNP检测方法  36
      3.2 技术原理及优点  36-39
    4 湖羊起源与品种特点及多胎研究现状  39-43
      4.1 湖羊起源研究  39-40
      4.2 湖羊的品种特点  40-41
      4.3 湖羊多胎研究状况  41-43
第二部分试验研究  43-83
  第一章 FecB基因在7个绵羊品种中的多样性研究  43-52
    1 材料与方法  43-46
      1.1 样本采集与保存  43-44
      1.2 主要仪器、设备及试剂材料  44
      1.3 样本DNA提取  44-45
      1.4 DNA纯度检测  45
      1.5 FecB基因多态性检测  45
      1.6 序列测定  45-46
      1.7 数据收集与统计分析  46
    2.结果  46-49
      2.1 基因组完整性  46
      2.2 不同群体湖羊的多产性  46-47
      2.3 FecB基因多样性  47-49
      2.4 FecB基因测序  49
    3.讨论  49-52
  第二章 BMP15基因在7个绵羊品种中的多样性研究  52-59
    1 材料与方法  52-54
      1.1 样本采集  52
      1.2 主要仪器、设备和试剂  52
      1.3 样本DNA提取与纯度检测  52-53
      1.4 BMP15基因检测  53-54
        1.4.1 BMP15基因FecX~H突变检测  53
        1.4.2 BMP15基因FecX~I突变检测  53
        1.4.3 BMP15基因FecX~G突变检测  53
        1.4.4 BMP15基因FecX~B突变检测  53-54
    2.结果  54-56
      2.1 FecX~X突变  54
      2.2 FecX~I突变  54-55
      2.3 FecX~B突变  55-56
      2.4 FecX~G突变  56
    3.讨论  56-59
  第三章 GDF9基因在7个绵羊品种中的多样性研究  59-63
    1 材料与方法  59-60
      1.1 样本采集  59
      1.2 主要仪器、设备和试剂  59
      1.3 样本DNA提取与纯度检测  59
      1.4 GDF9基因FecG~H多态性检测  59-60
    2.结果  60-61
    3.讨论  61-63
  第四章湖羊微卫星多态性研究  63-76
    1 材料与方法  63-67
      1.1 样本采集  63-64
      1.2 微卫星标记的选择  64
      1.3 主要仪器、设备和试剂  64
      1.4 样本DNA提取和检测  64
      1.5 PCR扩增  64-65
      1.6 电泳结果记录及处理  65
      1.7 数据统计及分析  65-67
    2 结果  67-72
      2.1 基因型检测  67
      2.2 微卫星位点等位基因频率分布  67-69
      2.3 微卫星基因型(频率)与湖羊产羔数的关系  69-70
      2.4 品种内的遗传变异  70-72
      2.5 群体间遗传距离和相似系数  72
    3 讨论  72-76
  第五章 FecB基因检测新方法(Tetra-ARMS PCR)建立  76-83
    1.材料与方法  76-77
      1.1 实验动物与样本采集  76
      1.2 仪器与试剂材料  76-77
      1.3 基因组DNA提取  77
    2.FecB基因检测及Tetra-ARMS PCR引物设计  77-79
      2.1 FecB基因PCR-RFLP检测  77
      2.2 Tetra-ARMS PCR特异性引物设计  77-78
      2.3 参照引物设计  78
      2.4 四引物ARMS PCR扩增体系优化  78
      2.5 四引物ARMS PCR扩增和电泳  78-79
      2.6 四引物ARMS PCR引物扩增结果判读和意义  79
    3.结果  79-80
      3.1 四引物ARMS PCR扩增条件优化  79
      3.2 引入错配对特异性的影响  79-80
      3.3与PCR-RFLP方法的比较  80
    4.讨论  80-83
参考文献  83-93
全文结论  93-95
创新  95-96
附录  96-101
博士在读期间发表论文情况  101-103
致谢  103

绵羊多产性状的DNA分子标记及相关研究

内容摘要

全文目录

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