LIF基因多态性检测及其与母猪产仔性状的关联性分析

发布时间：2018-06-09 05:15

本文选题：猪 + LIF基因　；参考：《安徽农业大学》2015年硕士论文

【摘要】：本研究以安徽省优良地方猪种(安庆六白猪、圩猪和皖南黑猪)和外来瘦肉型猪种大白猪为试验材料,研究动物胚胎附植相关候选基因白血病抑制因子基因(leukemia inhibitory factor,LIF)的单链核苷酸多态性(single nucleotide polymerphisms,SNPs)检测及其与繁殖母猪产仔性状的相关性。根据NCBI中公布的猪LIF(登录号:AJ296176)基因外显子序列设计引物,采用PCR-SSCP、PCR-RFLP和PCR产物双向测序技术检测LIF基因第2、3外显子部分片段的遗传变异,筛查SNPs并分析其多态性,运用SPSS程序22.0 GLM模型分析遗传变异与繁殖母猪产仔性状的相关性,以探讨利用该基因作为分子标记改良母猪产仔性状的可行性,为安徽地方猪种繁殖性状的遗传改良提供科学根据。试验结果如下:(1)LIF基因外显子2扩增片段在安庆六白猪群体发现1个SNP,为LIFg.C5709T;外显子3扩增片段在安庆六白猪、圩猪和大白猪群体发现1个SNP,为LIFg.C6646T、在圩猪和大白猪群体发现1个SNP,为LIFg.C6988T。(2)群体遗传学分析结果表明:LIFg.C5709T在安庆六白猪群体中只检测到2种基因型(CC、CT),其中CC和CT基因型频率分别为70.71%、29.29%,该位点在安庆六白猪群体的多态信息含量(PIC)为0.2188,处于低度多态,卡方适合性检验p值大于0.05,处于Hardy-Weinberg平衡状态;LIFg.C6646T在安庆六白猪、圩猪和大白猪群体中均检测到3种基因型(AA、AB和BB),其中AA基因型频率在3个群体中分别为47.47%、57.14%和50.62%,AB基因型频率分别为42.42%、35.16%、35.80%,BB基因型频率分别为10.11%、7.70%、13.58%,该位点在3个群体的多态信息含量(PIC)分别为0.3377、0.3064、0.3383,在该位点均处于中度多态,卡方适合性检验p值均大于0.05,处于Hardy-Weinberg平衡状态;LIFg.C6988T在圩猪和大白猪群体中均检测到3种基因型(DD、DE和EE),其中DD基因型频率在这2个群体中分别为45.05%、45.91%,DE基因型频率分别为32.97%、39.51%,EE基因型频率分别为21.98%、13.58%,该位点在圩猪和大白猪群体的多态信息含量(PIC)分别为0.3613、0.3457,均处于中度多态,卡方适合性检验p值均大于0.05,处于Hardy-Weinberg平衡状态。(3)LIF基因突变位点与繁殖母猪产仔性状的相关性分析:LIFg.C5709T位点:基因型对安庆六白猪初产母猪和经产母猪总产仔数(TNB)和产活仔数(NBA)均无显著影响,但均存在CC型高于CT型的趋势。LIFg.C6646T位点:安庆六白猪初、经产母猪群体BB基因型的TBA和NBA均极显著高于AB和AA基因型(P0.01),初产母猪群体AB基因型极显著高于AA基因型(P0.01)。大白猪群体初产母猪AA基因型的TNB和NBA均极显著高于AB和BB基因型(P0.01);经产母猪AA基因型TNB显著高于AB基因型(P0.05),AA基因型NBA极显著高于AB基因型(P0.01)。LIFg.C6988T位点:圩猪初、经产母猪群体DD基因型TNB和NBA极显著高于DE和EE基因型(P0.01)。大白猪初产母猪群体DD基因型TNB和NBA极显著高于DE和EE基因型(P0.01);经产母猪DD基因型TNB极显著高于DE和EE基因(P0.01),DD基因型NBA极显著高于DE和EE基因型(P0.01),DE基因型NBA显著高于EE基因型。研究结果提示,LIFg.C6646T位点可以作为安庆六白猪、大白猪繁殖母猪产仔性状候选遗传标记,LIFg.C6988T位点可以作为圩猪、大白猪繁殖母猪产仔性状候选遗传标记,以应用于繁殖母猪产仔性状的改良。
[Abstract]:In this study, the single strand nucleoside polymorphism (single nucleotide polymerphisms, SNPs) of the candidate gene leukemia inhibitory factor (LIF) gene (leukemia inhibitory factor, LIF) was detected by the experimental materials of the fine local pig breeds (six white pigs, polder pigs and southern Anhui pigs) and foreign lean pig breeds. According to the correlation between the litter size and the reproductive performance of the breeding sows, the primers were designed according to the exon sequence of the pig LIF (login number: AJ296176) published in NCBI. The genetic variation of the partial fragment of the exon 2,3 of the LIF gene was detected by PCR-SSCP, PCR-RFLP and PCR products, and the polymorphism of the LIF gene was screened and analyzed. The SPSS program 22 GLM module was used. The correlation between genetic variation and reproductive sow litter traits was analyzed in order to explore the feasibility of using this gene as a molecular marker to improve the litter size of sows and to provide scientific basis for genetic improvement of Anhui local pig breeding traits. The results were as follows: (1) 1 SNP of LIF gene exon 2 amplified fragment was found in six white pig population in Anqing. For LIFg.C5709T, exon 3 amplified fragments in Anqing six white pigs, polder pigs and big white pigs, found 1 SNP, LIFg.C6646T, 1 SNP in polder and white pig populations, and LIFg.C6988T. (2) population genetics analysis showed that only 2 genotypes (CC, CT) were detected in LIFg.C5709T in Anqing six white pigs population, CC and CT genotype frequency. The rate was 70.71%, 29.29%, the polymorphism information content (PIC) of the six white pig population in Anqing was 0.2188, in low polymorphism, the p value of chi square fitness test was more than 0.05, in the state of Hardy-Weinberg equilibrium; 3 genotypes (AA, AB and BB) were detected in the Anqing six white pig, the polder pig and the big white pig population, in which the AA genotype frequency was the frequency. The rate was 47.47%, 57.14% and 50.62% in 3 groups, and the genotype frequencies of AB were 42.42%, 35.16% and 35.80%, respectively, and the genotype frequencies were 10.11%, 7.70%, 13.58%, respectively, and the polymorphism information content (PIC) was 0.3377,0.3064,0.3383 in 3 groups, respectively, and the p value of chi square fitness test was greater than 0.05, and the p value of chi square was greater than 0.05. In the equilibrium state of Hardy-Weinberg, 3 genotypes (DD, DE and EE) were detected in the polder and white pig populations, in which the frequency of DD genotype was 45.05% and 45.91% in the 2 populations, and the frequencies of the DE genotype were 32.97%, 39.51%, and 21.98%, 13.58%, respectively, and the polymorphism of the loci was polymorphic in the polder and white pig populations. The information content (PIC) was 0.3613,0.3457, all in moderate polymorphism, the chi square fitness test p value was more than 0.05, in the Hardy-Weinberg equilibrium state. (3) the correlation analysis between the LIF gene mutation site and the reproductive sow litter character: LIFg.C5709T locus: the genotype to the total number (TNB) and production of the Anqing six white pigs and the production sows. There was no significant effect on the number of live litter (NBA), but there was a trend.LIFg.C6646T locus of type CC higher than that of CT type: the TBA and NBA of BB genotypes of the Anqing six white pigs were significantly higher than that of AB and AA genotypes (P0.01), and the AB genotypes of the first sow population were significantly higher than those of the AA genotypes. And NBA was significantly higher than that of AB and BB genotypes (P0.01), AA genotype TNB was significantly higher than AB genotype (P0.05), and AA genotype NBA was significantly higher than AB genotype (P0.01) locus. And NBA was significantly higher than DE and EE genotype (P0.01), and DD genotype TNB was significantly higher than DE and EE gene (P0.01), and DD genotype NBA was significantly higher than DE and genotype. Genetic markers and LIFg.C6988T loci can be used as candidate genetic markers for litter size traits in the breeding pigs of Wei pig and large white pig, so as to improve the litter size of breeding sows.
【学位授予单位】：安徽农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S828

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