绵羊ADIPOQ、UCP1和FGF21基因遗传变异与生长及胴体肌肉性状关联性研究

发布时间：2018-05-29 07:45

  本文选题：绵羊 + ADIPOQ　； 参考：《甘肃农业大学》2016年博士论文

【摘要】：本研究以ADIPOQ,UCP1和FGF21基因为候选基因,应用PCR-SSCP技术,针对美利奴羊(Merino)、特克赛尔羊(Texel)、丘陵陶赛特羊(Dorset Down)、新西兰罗姆尼羊(New Zealand Romney)、萨福克羊(Suffolk)、考力代尔羊(Corriedale)、杜泊羊(Dorper)、派伦代羊(Perendale)和藏绵羊(Tibetan sheep)9个绵羊品种,探索了上述3个基因共16个区域的核苷酸序列。在对多态性较为丰富的基因区域(如ADIPOQ基因外显子1,UCP1基因启动子区和FGF21基因外显子3区),用来自不同公羊家系1200只左右新西兰罗姆尼羔羊,分析核苷酸变异及单体型对生长性状和胴体肌肉性状的影响,探讨候选基因遗传变异作为绵羊生长、脂肪和胴体性状分子标记的可能性。主要研究结果如下:1.ADIPOQ,UCP1和FGF21基因多态性(1)在ADIPOQ基因启动子区检测到4条变异序列(A1-D1)和7个SNPs位点;在外显子2区检测到4条变异序列(A2-D2)和6个SNPs位点;在外显子3-1区检测到3条变异序列(A3-C3)和3个SNPs位点;在外显子3-2区和3’UTR区均检测到2条变异序列(A4-B4和A5-B5),分别包含2个SNPs位点和1个SNPs位点。在所检测到的19个SNPs位点中,有2个位于编码区的变异且为非同义突变,分别为位于第2外显子的c.46 T/C突变导致p.Tyr16His氨基酸变异,位于第3外显子的c.515 A/G导致p.Lys172Arg氨基酸变异。(2)在UCP1基因启动子1区共检测到6条变异序列(A1-F1)和12个SNPs位点;在启动子2区共检测到3条变异序列(A2-C2)和2个SNPs位点;在内含子1-1区共检测到5条变异序列(A3-E3)和8个SNPs位点;在内含子1-2区没有检测到多态位点;在内含子5区共检测到3条变异序列(A5-C5)和6个SNPs位点;在外显子6区共检测到3条变异序列(A6-C6)和2个SNPs位点;在所检测到的30个SNPs位点中,有1个在编码区的变异为非同义突变,为位于第6外显子的c.910 G/A突变导致p.Ala304Thr氨基酸变异。(3)在FGF21基因启动子1-外显子1区检测到2条变异序列(A1-B1)和2个SNPs位点;在外显子1-外显子2区检测到4条变异序列(A2-D2)和6个SNPs位点;在外显子2区检测到2条变异序列(A3-B3)和7个SNPs位点;在内含子2区检测到3条变异序列(A4-C4),包含3个SNPs位点:在内含子2-3’UTR区检测到4条变异序列(A5-D5),包含4个SNPs位点。在所检测到的22个SNPs位点中,有2个在编码区的变异为非同义突变,分别为位于第1外显子的c.161 T/C突变导致p.Ala54Val氨基酸变异,位于第3外显子的c.554 C/T导致p.Pro185Leu氨基酸变异。2.ADIPOQ,UCP1和FGF21基因突变及单体型对新西兰罗姆尼绵羊生长性状的影响(1)等位基因分析结果表明,ADIPOQ基因启动子区等位基因只与断尾时体重关联(P0.05),对其它生长性状无影响(P0.05),且缺失等位基因A1的个体有较高的断尾重。第2内含子等位基因对所有生长性状无影响(P0.05)。单体型分析表明,由启动子区和内含子2区构成的8种单体型中,单体型A1-A3与断奶前平均生长速度关联(P0.05),缺失单体型A1-A3的群体有更高的生长速度。(2)等位基因分析结果表明,UCP1基因启动子区等位基因对所有生长性状无影响(P0.05)。第5内含子等位基因与初生重关联(P0.05),存在等位基因C5的群体初生重高于缺失群体。单体型分析结果表明,单体型与断奶前各生长性状无显著相关性(P0.05)。(3)等位基因分析结果表明,FGF21基因外显子3区等位基因对生产性状无影响(P0.05),但基因型对断奶前平均生长速度有显著影响(P0.05),且存在基因型A5A5的群体较其它基因型有更快的生长速度。3.ADIPOQ,UCP1和FGF21基因突变及单体型对新西兰罗姆尼羊胴体肌肉性状的影响(1)ADIPOQ基因启动子区等位基因A1与热胴体重、后腿瘦肉量、腰部瘦肉量和总瘦肉量存在关联性(P0.05),等位基因B1与后腿瘦肉量、后腿瘦肉比例和肩部瘦肉量存在关联性(P0.05),其它胴体性状与等位基因无关联性;基因型与热胴体重、后腿瘦肉量、腰部瘦肉量和总瘦肉量有关联性(P0.05)。第2内含子等位基因C3与GR值有关联性(P0.05),其它胴体性状与等位基因无关联性(P0.05);基因型与GR值有关联性(P0.05)。单体型A1-A3与后腿瘦肉量、腰部瘦肉量和总瘦肉量有关联性(P0.05),A1-C3与GR值和腰部瘦肉比例有关联性(P0.05),B1-A3与后退瘦肉比例和肩部瘦肉比例有关联性(P0.05)。双体型与热胴体重、后腿瘦肉量、腰部瘦肉量和总瘦肉量有关联性(P0.05)。(2)UCP1基因启动子区等位基因B2与热胴体重关联(P0.05),其它等位基因与胴体无关联性(P0.05);基因型也只与热胴体重有关联性(P0.05)。第5内含子区等位基因与基因型与各胴体肌肉性状均无关联性(P0.05)。单体型A2-B5与热胴体重和腰部瘦肉量有关联性(P0.05),单体型C2-C5与后腿瘦肉比例有关联性(P0.05),对其它胴体肌肉性状无影响。(3)FGF21基因各区域变异位点之间存在很高的连锁关系,因此只对高度多态的内含子2-外显子3区与绵羊生长于胴体肌肉性状进行关联性分析,结果表明,等位基因均对各胴体肌肉性状无显著关联影响(0.05P),只有等位基因A5对GR值有一定的影响趋势(0.05P0.2),各基因型均对胴体肌肉性状无影响(0.05P)。综上所述,ADIPOQ基因、UCP1基因和FGF21基因均具有丰富的多态性,且ADIPOQ基因和UCP1基因的部分等位基因及单体型可作为绵羊生长及胴体肌肉性状的候选分子育种标记。
[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.
【学位授予单位】：甘肃农业大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：S826
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本文编号：1950066