嘉兴黑猪繁殖性状相关基因ESR、FSHβ、PRLR多态性及其关联分析研究

发布时间：2018-05-21 10:16

本文选题：嘉兴黑猪 + 雌激素受体　；参考：《浙江农林大学》2015年硕士论文

【摘要】：猪繁殖性状被公认为是低遗传力的数量性状和经济性状,中国地方猪种在这一性状表现比较优良。在猪遗传育种过程中,传统杂交育种和分子标记辅助选择育种相结合(marker-assisted selection,MAS)成为现代选种、保种技术的新方向。本研究中以嘉兴黑猪为研究对象,选取雌激素受体(Estrogen Receptor,ESR)基因、卵泡刺激素β亚基(Follicle-Stimulating Hormoneβ,FSHβ)基因、催乳素受体(Prolactin Receptor,PRLR)基因为候选基因,采取513份嘉兴黑猪耳组织为试验材料,抽提其基因组DNA,以候选基因的外显子区域设计引物,PCR扩增得到目的序列,目的片段采用PCR-SSCP方法和直接测序方法检测SNPs,统计分析各基因的等位基因频率和基因型频率、Hardy-Weinberg平衡情况以及其与繁殖性状关联分析结果,为建立嘉兴黑猪的分子标记辅助育种模型奠定基础。ESR基因第一外显子区域扩增目的片段,测序后分析得到4个SNPs:T6901C、G6759A、T6927G、A7188G。测序峰图表明每个SNP处都出现三种基因型,分别命名为AA、AB、BB、NN、NM、MM、EE、EF、FF、JJ、HJ和HH。统计分析了这些位点的基因型频率和基因频率,卡方检验表明,T6901C、A7188G两个基因座均处于Hardy-Weinberg平衡状态,G6759A、T6927G显著偏离Hardy-Weinberg平衡状态。选取嘉兴黑猪产仔数、活仔数、死胎数、胎间距、初生均重5个繁殖相关性状与多态性位点T6901C、A7188G的六种基因型分别进行性状关联分析结果表明,T6901C位点与性状关联分析三个基因型之间无显著性差异;而A7188G位点中HH基因型个体产仔数与活仔数均显著高于JJ基因型个体(P0.05),死胎数性状HH基因型显著高于JJ基因型(P0.05),初生重性状HH性状显著高于JJ基因型(P0.05),在其他性状,各基因型之间无显著性差异。FSHβ基因的外显子2上扩增的目的片段测序分析获得到3个SNPs:T1035C、A922C、A925G。每个SNP位点都出现三种基因型,分别命名为KK、KL、LL、QQ、QR、RR、UU、UV、VV。统计分析了这些位点的基因频率和基因型频率,卡方检验表明,A922C位点显著偏离Hardy-Weinberg平衡状态,T1035C、A925G位点符合Hardy-Weinberg平衡状态。T1035C、A925G位点各基因型与嘉兴黑猪群体5个繁殖性状关联分析,结果显示T1035C位点各基因型之间无显著性差异,G925A位点分析发现,UU基因型与VV基因型个体的初生重显著高于UV基因型(P0.05)。PRLR基因第5外显子区域发现1个SNPs:A13902T,PCR-SSCP检测显示存在三种基因型:XX、XY和YY。统计分析了这些位点的基因频率和基因型频率,卡方检验表明,此位点符合Hardy-Weinberg平衡状态;与5个繁殖性状关联分析结果显示,产仔数性状、活仔数性状、初生重性状在A13902T位点的基因型之间差异显著(P0.01)XXXYYY,与其他性状各基因型之间则无显著性差异。A7188G位点与A13902T两个位点合并基因型效应对于五个繁殖性状关联分析发现,合并基因型中,产仔数性状HHXX基因型比JJXX基因型每胎多产2.03头(P0.05),HJXX基因型比JJXY基因型每胎多产2.12头(P0.05),比JJYY基因型每胎多产2.38头(P0.05)。对于活仔数性状HHXX基因型比JJXX基因型每胎多1.95头(P0.05),比JJYY基因型多3.53头(P0.03),初生重性状HHXX基因型每窝比JJYY基因型重4.37kg(P0.05),比JJXY基因型每窝重3.83kg(P0.05)。死胎数性状与胎间距性状没有显著影响。
[Abstract]:The reproductive traits of pigs are considered to be quantitative and economic traits of low heritability, and Chinese local pigs have excellent performance in this character. In the process of swine genetics and breeding, the combination of traditional hybrid breeding and molecular marker assisted selection breeding (marker-assisted selection, MAS) has become a modern selection and the new direction of seed conservation. Taking the Jiaxing black pig as the research object, we selected the estrogen receptor (Estrogen Receptor, ESR) gene, the follicle stimulating hormone beta subunit (Follicle-Stimulating Hormone beta, FSH beta) gene, the prolactin receptor (Prolactin Receptor, PRLR) gene as the candidate gene, and took 513 copies of the Jiaxing black pig ear tissue as the test material, and extracted its genome DNA, which was a candidate base. The exons were designed with primers, and the target sequence was amplified by PCR. The target fragment was detected by PCR-SSCP method and direct sequencing method for SNPs. The allele frequency and genotype frequency of each gene, Hardy-Weinberg balance and its correlation with reproductive traits were analyzed to assist the establishment of molecular markers in Jiaxing black pig. The breeding model laid the base of the first exon of.ESR gene to amplify the target fragment. After sequencing, 4 SNPs:T6901C, G6759A, T6927G, and A7188G. sequencing peak maps showed that there were three genotypes at each SNP, which were named AA, AB, BB, NN, NM. Rate, chi square test showed that the two loci of T6901C and A7188G were all in the state of Hardy-Weinberg balance, G6759A and T6927G deviated from the equilibrium state of Hardy-Weinberg significantly. The number of litter size, the number of live births, the number of stillbirths, the distance between the fetus, and the primary weight of the Jiaxing black pigs were all 5 reproductive correlation and polymorphic loci T6901C, and the six genotypes of A7188G were respectively carried out. The correlation analysis showed that there was no significant difference between the three genotypes of the T6901C loci and the traits, while the number of litter and the live number of the HH genotype in the A7188G locus were significantly higher than that of the JJ genotype (P0.05). The HH genotype of the number of stillbirths was significantly higher than that of the JJ genotype (P0.05), and the HH traits of the primary weight traits were significantly higher than the JJ genotype (P). 0.05), in other traits, there was no significant difference between the genotypes, and the sequence analysis of exon 2 of.FSH beta gene was sequenced. Three genotypes were found at 3 SNPs:T1035C, A922C, and A925G. at each SNP site, named KK, KL, LL, QQ, QR, RR, UU, and genotype frequencies. The chi square test showed that the A922C locus was significantly deviated from the equilibrium state of Hardy-Weinberg, T1035C and A925G were in accordance with Hardy-Weinberg equilibrium state.T1035C, and the genotype of A925G loci was associated with 5 reproductive traits in Jiaxing black pig population. The results showed no significant difference between the genotypes of the T1035C loci, the G925A site analysis found, the UU genotypes and the genotype of the.T1035C. The primary weight of the VV genotype was significantly higher than that of the UV genotype (P0.05).PRLR gene fifth exon region found 1 SNPs:A13902T, PCR-SSCP detection showed three genotypes: XX, XY and YY. statistical analysis of the gene frequency and genotype frequency of these loci, chi square test showed that this site was in line with Hardy-Weinberg equilibrium state; and 5 propagation. The results of the correlation analysis showed that the number of litter, the number of live litter, and the genotypes of the primary weight traits were significantly different (P0.01) XXXYYY, and there was no significant difference between the genotypes of the other traits and the other traits. The combination of the genotype effects of the.A7188G and A13902T two sites was found to be found in the association analysis of the two reproductive traits. In the genotype, the number of HHXX genotypes was 2.03 more (P0.05) than that of the JJXX genotype, and the HJXX genotype was 2.12 more (P0.05) per fetus than the JJXY genotype, and 2.38 more (P0.05) more than the JJYY genotype per tire. The HHXX genotype of the living litter was 1.95 more (P0.05) than the JJXX genotype, and 3.53 more than the JJYY genotype. The weight per litter of HHXX genotype was more than JJYY genotype 4.37kg (P0.05), and 3.83kg (P0.05) per litter weight of JJXY genotypes. There was no significant effect on the number of stillbirths and fetal distance traits.
【学位授予单位】：浙江农林大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S828

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