【作者】 徐珊珊；

【导师】 蒋思文；

【作者基本信息】 华中农业大学 ， 动物遗传育种与繁殖， 2008， 硕士

【摘要】 繁殖性状是非常重要的经济性状,产仔数的高低直接影响养猪业的生产效益,而排卵率和产仔数呈正相关。根据基因的生理生化功能,本研究选择了影响排卵率的候选基因含凝血酶敏感蛋白模体的去解联金属蛋白酶(a disintegrin andmetalloproteinase with thrombospondin motifs-1,ADAMTS-1),应用比较基因组学和生物信息学等方法分离鉴定候选基因;利用直接测序法对所分离片段进行SNP筛选;利用PCR-RFLP方法对SNP进行检测,并以长白猪、大白猪、杜洛克原种猪群为材料,研究了猪ADAMTS-1基因的多态性位点与猪产仔数的相关性。具体结果如下:1.获得长白猪、大白猪、杜洛克猪ADAMTS-1基因的包括第六、七外显子及第六内含子部分基因组序列,经比对发现该基因外显子7编码区72bp处C/G颠换的SNP导致氨基酸改变(精氨酸变为脯氨酸),导致了PvuⅡ限制性内切酶长度多态性,据此建立了PCR-PvuⅡ-RFLP酶切分型方法。2.在长白猪、大白猪、杜洛克猪群中采用PCR-PvuⅡ-RFLP酶切分型技术,进行ADAMTS-1基因的多态性检测,并与繁殖性状进行了关联分析,结果表明,在长白猪、大白猪、杜洛克猪中,均存在AA,AB,BB三种基因型。在长白猪中,初产的AA基因型产仔数最高,死胎数最低,BB型的死胎数最高;经产的产仔和活产仔数均是BB＞AB＞AA,差异显著(p＜0.05),且加性效应显著(p＜0.05)。在大白猪中初产的产仔数和活仔数均为AB型最高;经产的产仔数和活仔数均为BB型最高。在杜洛克猪中初产和经产的AB基因型的产仔数和活仔数均高于AA基因型和BB基因型。3.获得长白猪、大白猪、杜洛克猪ADAMTS-1基因的包括第七、八外显子及第七内含子部分基因组序列,经比对发现该基因第七内含子926bp处有一个A/G转换,建立了PCR-MvaⅠ-RFLP方法酶切分型方法。4.在长白猪、大白猪、杜洛克猪群中采用PCR-MvaⅠ-RFLP酶切分型技术,进行ADAMTS-1基因的多态性检测,并与繁殖性状进行了关联分析。结果表明,在长白猪、大白猪、杜洛克猪中,均存在AA,AG,GG三种基因型,但是A等位基因频率在三种猪中明显偏低。在长白猪中,AG基因型的繁殖性状优于GG,在经产大白猪和初产杜洛克猪中AA基因型的产仔性状优于AG和GG基因型,推测A是优势等位基因,仍需进一步验证。更多还原

【Abstract】 The economic efficiency of swine industry is influenced by litter size deeply and there was a positive correlation between ovulation rate and litter size.According to physiological or biochemical functions of genes,selected the candidate gene ADAMTS-1(a disintegrin and metalloproteinase with thrombospondin motifs-1),we isolated and identified ADAMTS-1 with comparative genomics and bioinformatic approaches, found some SNPs by direct sequencing and used PCR-RFLP detected these SNPs. Landrace,Large White,Duroc pig breeds were used to study the relationship of the polymorphism of the genes and association studies of the gene polymorphism with reproduction traits.The main results are as following:1.The nucleotide sequences in exon7 of ADAMTS-1 gene of Landrace,Large White,Duroc pig breeds were obtained with cloning and sequencing. Comparative sequencing of the amplified fragments revealed an PCR-PvuⅡ-RFLP was developed to detect the C/G substitution in exon7 which resulted in amino acid change(Arg→Pro).2.From the description of the frequencies of genotype and alleles in three groups, three genotypes were detected,and do association analysis with reproduction traits.Three genotypes(AA,AB,BB)existed in these breeds.Results showed that there was significantly difference between genotype AA and BB with LS and NBA for primiparous and multiparous in Landrace breed (p＜0.05),the additive effect is significant(p＜0.05).Also there was difference between three genotypes in Large White and Duroc pig breeds3.PCR- MvaⅠ-RFLP was developed to scan another SNP which didn’t result in amino acid change.In intron 7 of ADAMTS-1 gene,comparative sequencing of the amplified fragments and find an A/G substitution.4.From the description of the frequencies of genotype and alleles in Landrace, Large White,Duroc pig groups,three genotypes were detected,and do association analysis with reproduction traits.Three genotypes(GG,GB, BB)existed in these breeds.Also there was difference between three genotypes in Landrace,Large White and Duroc pig breeds.The frequency of the allele G was low,but the reproductive traits of the allele G were excellent. We Conjectured the allele G maybe the preponderant alleles,need further verification.更多还原