發(fā)布時(shí)間：2018-06-09 05:15

  本文選題：豬 + LIF基因��； 參考：《安徽農(nóng)業(yè)大學(xué)》2015年碩士論文

【摘要】：本研究以安徽省優(yōu)良地方豬種(安慶六白豬、圩豬和皖南黑豬)和外來瘦肉型豬種大白豬為試驗(yàn)材料,研究動(dòng)物胚胎附植相關(guān)候選基因白血病抑制因子基因(leukemia inhibitory factor,LIF)的單鏈核苷酸多態(tài)性(single nucleotide polymerphisms,SNPs)檢測及其與繁殖母豬產(chǎn)仔性狀的相關(guān)性。根據(jù)NCBI中公布的豬LIF(登錄號(hào):AJ296176)基因外顯子序列設(shè)計(jì)引物,采用PCR-SSCP、PCR-RFLP和PCR產(chǎn)物雙向測序技術(shù)檢測LIF基因第2、3外顯子部分片段的遺傳變異,篩查SNPs并分析其多態(tài)性,運(yùn)用SPSS程序22.0 GLM模型分析遺傳變異與繁殖母豬產(chǎn)仔性狀的相關(guān)性,以探討利用該基因作為分子標(biāo)記改良母豬產(chǎn)仔性狀的可行性,為安徽地方豬種繁殖性狀的遺傳改良提供科學(xué)根據(jù)。試驗(yàn)結(jié)果如下:(1)LIF基因外顯子2擴(kuò)增片段在安慶六白豬群體發(fā)現(xiàn)1個(gè)SNP,為LIFg.C5709T;外顯子3擴(kuò)增片段在安慶六白豬、圩豬和大白豬群體發(fā)現(xiàn)1個(gè)SNP,為LIFg.C6646T、在圩豬和大白豬群體發(fā)現(xiàn)1個(gè)SNP,為LIFg.C6988T。(2)群體遺傳學(xué)分析結(jié)果表明:LIFg.C5709T在安慶六白豬群體中只檢測到2種基因型(CC、CT),其中CC和CT基因型頻率分別為70.71%、29.29%,該位點(diǎn)在安慶六白豬群體的多態(tài)信息含量(PIC)為0.2188,處于低度多態(tài),卡方適合性檢驗(yàn)p值大于0.05,處于Hardy-Weinberg平衡狀態(tài);LIFg.C6646T在安慶六白豬、圩豬和大白豬群體中均檢測到3種基因型(AA、AB和BB),其中AA基因型頻率在3個(gè)群體中分別為47.47%、57.14%和50.62%,AB基因型頻率分別為42.42%、35.16%、35.80%,BB基因型頻率分別為10.11%、7.70%、13.58%,該位點(diǎn)在3個(gè)群體的多態(tài)信息含量(PIC)分別為0.3377、0.3064、0.3383,在該位點(diǎn)均處于中度多態(tài),卡方適合性檢驗(yàn)p值均大于0.05,處于Hardy-Weinberg平衡狀態(tài);LIFg.C6988T在圩豬和大白豬群體中均檢測到3種基因型(DD、DE和EE),其中DD基因型頻率在這2個(gè)群體中分別為45.05%、45.91%,DE基因型頻率分別為32.97%、39.51%,EE基因型頻率分別為21.98%、13.58%,該位點(diǎn)在圩豬和大白豬群體的多態(tài)信息含量(PIC)分別為0.3613、0.3457,均處于中度多態(tài),卡方適合性檢驗(yàn)p值均大于0.05,處于Hardy-Weinberg平衡狀態(tài)。(3)LIF基因突變位點(diǎn)與繁殖母豬產(chǎn)仔性狀的相關(guān)性分析:LIFg.C5709T位點(diǎn):基因型對(duì)安慶六白豬初產(chǎn)母豬和經(jīng)產(chǎn)母豬總產(chǎn)仔數(shù)(TNB)和產(chǎn)活仔數(shù)(NBA)均無顯著影響,但均存在CC型高于CT型的趨勢。LIFg.C6646T位點(diǎn):安慶六白豬初、經(jīng)產(chǎn)母豬群體BB基因型的TBA和NBA均極顯著高于AB和AA基因型(P0.01),初產(chǎn)母豬群體AB基因型極顯著高于AA基因型(P0.01)。大白豬群體初產(chǎn)母豬AA基因型的TNB和NBA均極顯著高于AB和BB基因型(P0.01);經(jīng)產(chǎn)母豬AA基因型TNB顯著高于AB基因型(P0.05),AA基因型NBA極顯著高于AB基因型(P0.01)。LIFg.C6988T位點(diǎn):圩豬初、經(jīng)產(chǎn)母豬群體DD基因型TNB和NBA極顯著高于DE和EE基因型(P0.01)。大白豬初產(chǎn)母豬群體DD基因型TNB和NBA極顯著高于DE和EE基因型(P0.01);經(jīng)產(chǎn)母豬DD基因型TNB極顯著高于DE和EE基因(P0.01),DD基因型NBA極顯著高于DE和EE基因型(P0.01),DE基因型NBA顯著高于EE基因型。研究結(jié)果提示,LIFg.C6646T位點(diǎn)可以作為安慶六白豬、大白豬繁殖母豬產(chǎn)仔性狀候選遺傳標(biāo)記,LIFg.C6988T位點(diǎn)可以作為圩豬、大白豬繁殖母豬產(chǎn)仔性狀候選遺傳標(biāo)記,以應(yīng)用于繁殖母豬產(chǎn)仔性狀的改良。
[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.
【學(xué)位授予單位】：安徽農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S828

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