本文選題：綿羊 + ADIPOQ�。� 參考：《甘肅農(nóng)業(yè)大學(xué)》2016年博士論文

【摘要】：本研究以ADIPOQ,UCP1和FGF21基因?yàn)楹蜻x基因,應(yīng)用PCR-SSCP技術(shù),針對(duì)美利奴羊(Merino)、特克賽爾羊(Texel)、丘陵陶賽特羊(Dorset Down)、新西蘭羅姆尼羊(New Zealand Romney)、薩�？搜�(Suffolk)、考力代爾羊(Corriedale)、杜泊羊(Dorper)、派倫代羊(Perendale)和藏綿羊(Tibetan sheep)9個(gè)綿羊品種,探索了上述3個(gè)基因共16個(gè)區(qū)域的核苷酸序列。在對(duì)多態(tài)性較為豐富的基因區(qū)域(如ADIPOQ基因外顯子1,UCP1基因啟動(dòng)子區(qū)和FGF21基因外顯子3區(qū)),用來(lái)自不同公羊家系1200只左右新西蘭羅姆尼羔羊,分析核苷酸變異及單體型對(duì)生長(zhǎng)性狀和胴體肌肉性狀的影響,探討候選基因遺傳變異作為綿羊生長(zhǎng)、脂肪和胴體性狀分子標(biāo)記的可能性。主要研究結(jié)果如下:1.ADIPOQ,UCP1和FGF21基因多態(tài)性(1)在ADIPOQ基因啟動(dòng)子區(qū)檢測(cè)到4條變異序列(A1-D1)和7個(gè)SNPs位點(diǎn);在外顯子2區(qū)檢測(cè)到4條變異序列(A2-D2)和6個(gè)SNPs位點(diǎn);在外顯子3-1區(qū)檢測(cè)到3條變異序列(A3-C3)和3個(gè)SNPs位點(diǎn);在外顯子3-2區(qū)和3’UTR區(qū)均檢測(cè)到2條變異序列(A4-B4和A5-B5),分別包含2個(gè)SNPs位點(diǎn)和1個(gè)SNPs位點(diǎn)。在所檢測(cè)到的19個(gè)SNPs位點(diǎn)中,有2個(gè)位于編碼區(qū)的變異且為非同義突變,分別為位于第2外顯子的c.46 T/C突變導(dǎo)致p.Tyr16His氨基酸變異,位于第3外顯子的c.515 A/G導(dǎo)致p.Lys172Arg氨基酸變異。(2)在UCP1基因啟動(dòng)子1區(qū)共檢測(cè)到6條變異序列(A1-F1)和12個(gè)SNPs位點(diǎn);在啟動(dòng)子2區(qū)共檢測(cè)到3條變異序列(A2-C2)和2個(gè)SNPs位點(diǎn);在內(nèi)含子1-1區(qū)共檢測(cè)到5條變異序列(A3-E3)和8個(gè)SNPs位點(diǎn);在內(nèi)含子1-2區(qū)沒(méi)有檢測(cè)到多態(tài)位點(diǎn);在內(nèi)含子5區(qū)共檢測(cè)到3條變異序列(A5-C5)和6個(gè)SNPs位點(diǎn);在外顯子6區(qū)共檢測(cè)到3條變異序列(A6-C6)和2個(gè)SNPs位點(diǎn);在所檢測(cè)到的30個(gè)SNPs位點(diǎn)中,有1個(gè)在編碼區(qū)的變異為非同義突變,為位于第6外顯子的c.910 G/A突變導(dǎo)致p.Ala304Thr氨基酸變異。(3)在FGF21基因啟動(dòng)子1-外顯子1區(qū)檢測(cè)到2條變異序列(A1-B1)和2個(gè)SNPs位點(diǎn);在外顯子1-外顯子2區(qū)檢測(cè)到4條變異序列(A2-D2)和6個(gè)SNPs位點(diǎn);在外顯子2區(qū)檢測(cè)到2條變異序列(A3-B3)和7個(gè)SNPs位點(diǎn);在內(nèi)含子2區(qū)檢測(cè)到3條變異序列(A4-C4),包含3個(gè)SNPs位點(diǎn):在內(nèi)含子2-3’UTR區(qū)檢測(cè)到4條變異序列(A5-D5),包含4個(gè)SNPs位點(diǎn)。在所檢測(cè)到的22個(gè)SNPs位點(diǎn)中,有2個(gè)在編碼區(qū)的變異為非同義突變,分別為位于第1外顯子的c.161 T/C突變導(dǎo)致p.Ala54Val氨基酸變異,位于第3外顯子的c.554 C/T導(dǎo)致p.Pro185Leu氨基酸變異。2.ADIPOQ,UCP1和FGF21基因突變及單體型對(duì)新西蘭羅姆尼綿羊生長(zhǎng)性狀的影響(1)等位基因分析結(jié)果表明,ADIPOQ基因啟動(dòng)子區(qū)等位基因只與斷尾時(shí)體重關(guān)聯(lián)(P0.05),對(duì)其它生長(zhǎng)性狀無(wú)影響(P0.05),且缺失等位基因A1的個(gè)體有較高的斷尾重。第2內(nèi)含子等位基因?qū)λ猩L(zhǎng)性狀無(wú)影響(P0.05)。單體型分析表明,由啟動(dòng)子區(qū)和內(nèi)含子2區(qū)構(gòu)成的8種單體型中,單體型A1-A3與斷奶前平均生長(zhǎng)速度關(guān)聯(lián)(P0.05),缺失單體型A1-A3的群體有更高的生長(zhǎng)速度。(2)等位基因分析結(jié)果表明,UCP1基因啟動(dòng)子區(qū)等位基因?qū)λ猩L(zhǎng)性狀無(wú)影響(P0.05)。第5內(nèi)含子等位基因與初生重關(guān)聯(lián)(P0.05),存在等位基因C5的群體初生重高于缺失群體。單體型分析結(jié)果表明,單體型與斷奶前各生長(zhǎng)性狀無(wú)顯著相關(guān)性(P0.05)。(3)等位基因分析結(jié)果表明,FGF21基因外顯子3區(qū)等位基因?qū)ιa(chǎn)性狀無(wú)影響(P0.05),但基因型對(duì)斷奶前平均生長(zhǎng)速度有顯著影響(P0.05),且存在基因型A5A5的群體較其它基因型有更快的生長(zhǎng)速度。3.ADIPOQ,UCP1和FGF21基因突變及單體型對(duì)新西蘭羅姆尼羊胴體肌肉性狀的影響(1)ADIPOQ基因啟動(dòng)子區(qū)等位基因A1與熱胴體重、后腿瘦肉量、腰部瘦肉量和總瘦肉量存在關(guān)聯(lián)性(P0.05),等位基因B1與后腿瘦肉量、后腿瘦肉比例和肩部瘦肉量存在關(guān)聯(lián)性(P0.05),其它胴體性狀與等位基因無(wú)關(guān)聯(lián)性;基因型與熱胴體重、后腿瘦肉量、腰部瘦肉量和總瘦肉量有關(guān)聯(lián)性(P0.05)。第2內(nèi)含子等位基因C3與GR值有關(guān)聯(lián)性(P0.05),其它胴體性狀與等位基因無(wú)關(guān)聯(lián)性(P0.05);基因型與GR值有關(guān)聯(lián)性(P0.05)。單體型A1-A3與后腿瘦肉量、腰部瘦肉量和總瘦肉量有關(guān)聯(lián)性(P0.05),A1-C3與GR值和腰部瘦肉比例有關(guān)聯(lián)性(P0.05),B1-A3與后退瘦肉比例和肩部瘦肉比例有關(guān)聯(lián)性(P0.05)。雙體型與熱胴體重、后腿瘦肉量、腰部瘦肉量和總瘦肉量有關(guān)聯(lián)性(P0.05)。(2)UCP1基因啟動(dòng)子區(qū)等位基因B2與熱胴體重關(guān)聯(lián)(P0.05),其它等位基因與胴體無(wú)關(guān)聯(lián)性(P0.05);基因型也只與熱胴體重有關(guān)聯(lián)性(P0.05)。第5內(nèi)含子區(qū)等位基因與基因型與各胴體肌肉性狀均無(wú)關(guān)聯(lián)性(P0.05)。單體型A2-B5與熱胴體重和腰部瘦肉量有關(guān)聯(lián)性(P0.05),單體型C2-C5與后腿瘦肉比例有關(guān)聯(lián)性(P0.05),對(duì)其它胴體肌肉性狀無(wú)影響。(3)FGF21基因各區(qū)域變異位點(diǎn)之間存在很高的連鎖關(guān)系,因此只對(duì)高度多態(tài)的內(nèi)含子2-外顯子3區(qū)與綿羊生長(zhǎng)于胴體肌肉性狀進(jìn)行關(guān)聯(lián)性分析,結(jié)果表明,等位基因均對(duì)各胴體肌肉性狀無(wú)顯著關(guān)聯(lián)影響(0.05P),只有等位基因A5對(duì)GR值有一定的影響趨勢(shì)(0.05P0.2),各基因型均對(duì)胴體肌肉性狀無(wú)影響(0.05P)。綜上所述,ADIPOQ基因、UCP1基因和FGF21基因均具有豐富的多態(tài)性,且ADIPOQ基因和UCP1基因的部分等位基因及單體型可作為綿羊生長(zhǎng)及胴體肌肉性狀的候選分子育種標(biāo)記。
[Abstract]:In this study, ADIPOQ, UCP1 and FGF21 genes were used as candidate genes, and PCR-SSCP was applied to Merino sheep (Merino), tksle sheep (Texel), Dorset Down, New Zealand Romney (New Zealand Romney), Suffolk sheep (Suffolk), Kolo Dyer sheep, Du Boyang, polendai sheep and Zang Mian The sheep (Tibetan sheep) 9 sheep breeds have explored the nucleotide sequences of the above 3 genes in 16 regions. The polymorphic region (such as exon 1 of ADIPOQ gene, UCP1 gene promoter region and FGF21 gene exon 3) and 1200 New Zealand Romney lambs from different goats' families were used to analyze nucleotide variation. And the effects of haplotype on growth traits and carcass muscle traits, and the possibility of genetic variation of candidate genes as molecular markers for sheep growth, fat and carcass traits. The main results are as follows: 1.ADIPOQ, UCP1 and FGF21 gene polymorphism (1) detected 4 mutation sequences (A1-D1) and 7 SNPs loci in the promoter region of the ADIPOQ gene; 4 mutation sequences (A2-D2) and 6 SNPs loci were detected in exon 2, 3 variation sequences (A3-C3) and 3 SNPs loci were detected in exon 3-1, 2 mutation sequences (A4-B4 and A5-B5) were detected in exon 3-2 and 3 'UTR region, and 2 SNPs loci and 1 SNPs loci respectively. There were 2 bits in the 19 SNPs loci of the detected. The mutation of the coding region and a non synonymous mutation, c.46 T/C mutations located in exon second, resulting in p.Tyr16His amino acid variation, and c.515 A/G in exon third lead to p.Lys172Arg amino acid variation. (2) 6 variation sequences (A1-F1) and 12 SNPs loci were detected in the UCP1 gene promoter 1; 3 bands were detected in the promoter 2. Variation sequence (A2-C2) and 2 SNPs loci; 5 variation sequences (A3-E3) and 8 SNPs loci were detected in intron 1-1; 3 mutation sequences (A5-C5) and 6 SNPs loci were detected in the intron 1-2; 3 variation sequences (A6-C6) and 2 SNPs loci were detected in the exon 6 region; Of the 30 SNPs loci detected, 1 of the mutations in the coding region were non synonymous mutations, and the c.910 G/A mutation located in exon sixth led to the p.Ala304Thr amino acid variation. (3) 2 mutation sequences (A1-B1) and 2 SNPs loci were detected in the 1 region of the FGF21 gene promoter 1- exon 1, and 4 mutation sequences were detected in the exon 1- exon 2 (A2-D). 2) and 6 SNPs loci; 2 mutation sequences (A3-B3) and 7 SNPs loci were detected in exon 2; 3 mutation sequences (A4-C4) were detected in the intron 2, including 3 SNPs loci: 4 sequences (A5-D5) detected in the intron 2-3 'UTR region, including 4 SNPs sites. In the 22 SNPs loci detected, there were 2 variations in the coding region. For non synonymous mutations, c.161 T/C mutations located in exon first lead to p.Ala54Val amino acid variation, and c.554 C/T in exon third leads to p.Pro185Leu amino acid variation.2.ADIPOQ, UCP1 and FGF21 mutations and the effect of haplotype on the growth traits of New Zealand Romney sheep (1) allele analysis results show that ADIPOQ base Because the promoter region allele was only associated with weight loss at the end of the tail (P0.05), there was no effect on other growth traits (P0.05), and the individuals with missing allele A1 had higher tail weight. Second intron alleles had no effect on all growth traits (P0.05). Haplotype analysis showed that in the 8 types of haplotypes composed of the promoter region and the intron 2 region, the single somatotype analysis showed that the single somatotype analysis showed that the single somatotype analysis showed that the single somatotype analysis showed that the single somatotype analysis was the only one of the 8 haplotypes, which were composed of the promoter region and the intron. Somatotype A1-A3 was associated with the average growth rate before weaning (P0.05), and there was a higher growth rate for the population with the deletion of the haplotype A1-A3. (2) the allelic analysis of the UCP1 gene promoter region had no effect on all growth traits (P0.05). Fifth intron alleles were associated with primary weight Association (P0.05), and there was a group of alleles C5. The results of haplotype analysis showed that there was no significant correlation between the haplotype and the growth traits before weaning (P0.05). (3) the allele analysis showed that the allele of exon 3 of the gene had no effect on the production traits (P0.05), but the genotype had a significant influence on the average growth rate before weaning (P0.05), and there was a gene. The population of type A5A5 has faster growth rate than other genotypes,.3.ADIPOQ, UCP1 and FGF21 gene mutations and haplotype effects on the carcass muscle properties of New Zealand Romney sheep (1) the ADIPOQ gene promoter region allele A1 and hot carcass weight, lean meat quantity in the hind legs, lean meat quantity in the waist and total lean meat (P0.05), allele B1 and The weight of lean meat in the hind legs, the proportion of lean meat in the hind legs and the weight of shoulder lean meat (P0.05), the other carcass traits are not related to the allele; the genotype is associated with the body weight of the carcass, the lean weight of the hind legs, the lean meat quantity in the waist and the total lean meat (P0.05). The second intron allele C3 is associated with the GR value (P0.05), other carcass traits and alleles The genotype was associated with the GR value (P0.05). The haplotype A1-A3 was associated with the lean meat quantity of the hind legs, the lean meat quantity of the waist and the total lean meat (P0.05), A1-C3 and the GR value and the lean lean meat ratio (P0.05). The proportion of B1-A3 to the lean lean meat ratio and the shoulder lean meat ratio (P0.05). Bis somatotype and hot carcass weight and lean hind legs. Meat quantity, lean meat quantity of waist and total lean meat were associated (P0.05). (2) the allele B2 of UCP1 gene promoter was associated with hot carcass weight (P0.05), other alleles were not associated with carcass (P0.05), and genotype was only associated with the body weight of hot carcass (P0.05). There was no correlation between allele and genotype of the inner subregion and the carcass muscle properties of the gene type (P0.05). Sex (P0.05). Haplotype A2-B5 is associated with hot carcass weight and lean meat quantity in the waist (P0.05), haplotype C2-C5 and hind leg lean meat ratio (P0.05), and no influence on other carcass muscle traits. (3) there is a high linkage between the loci of the FGF21 gene, so only the highly polymorphic intron 2- exon 3 and sheep The correlation analysis of carcass muscle traits showed that alleles had no significant influence on the carcass muscle properties (0.05P), only allele A5 had a certain influence on GR value (0.05P0.2), and all genotypes had no effect on carcass muscle properties (0.05P). In conclusion, ADIPOQ, UCP1, and FGF21 genes were all above all. The partial alleles and haplotypes of ADIPOQ and UCP1 genes can be used as candidate molecular markers for the growth of sheep and carcass muscle traits.
【學(xué)位授予單位】：甘肅農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類(lèi)號(hào)】：S826
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本文編號(hào)：1950066