贵州白山羊TFAM和TFB2M基因表达、多态对肉质、屠宰性状的影响研究
发布时间：2018-01-19 04:21
本文关键词： 贵州白山羊 肉质性状 屠宰性能 TFAM基因 TFB2M基因 关联性分析 出处：《贵州大学》2015年硕士论文 论文类型：学位论文
【摘要】：本研究以贵州白山羊为研究对象,采用PCR-SSCP、DNA直接测序、实时荧光定量PCR、克隆等方法克隆得到TFAM基因的cDNA序列并利用生物信息学分析TFAM蛋白的结构和理化性质等内容,同时探索了TFAM和TFB2M基因的mRNA表达规律和多态性对贵州白山羊肉质性状和屠宰性能的影响,为开展贵州白山羊肉质、屠宰性能的候选基因相关研究,提高地方山羊羊肉品质,加快品种选育提供依据。试验获得如下主要成果:1.首次得到的贵州白山羊TFAM基因的cDNA序列,通过生物信息学软件分析TFAM蛋白的结构和理化性质。TFAM蛋白ORF长741 bp,编码246个氨基酸。其理论等电点PI为9.74表明TFAM是酸性的,同时,TFAM蛋白氨基酸序列不存在跨膜螺旋结构不属于膜蛋白或分泌蛋白。TFAM蛋白功能分析表明TFAM蛋白在细胞被膜发挥功能的可能性最大,数值为13.18;基因本体分类表明,其可能发挥类似激素作用的概率数值为17.538。贵州白山羊和绵羊的氨基酸序列同源性为98.8%。2.试验筛选到TFB2M基因两个SNPs位点,分别是C-429T和A17110C,两个位点分别出现三种基因型CC、CT和TT,AA、AC和CC。群体遗传变异分析表明两个位点杂合度均较高,多态信息含量均表现为中度多态(0.25PIC0.50)。χ2检验表明,该群体在两个位点均符合Hardy-Weinberg平衡状态。TFAM基因没有筛选到SNPs。C-429T关联性分析结果显示,C-429T位点的CC和CT基因型个体系水力显著高于TT基因型个体(P0.05);TT基因型个体骨重显著大于CT基因型个体(P0.05),暗示该SNPs位点可能对贵州白山羊的肉质及骨重有影响。A17110C位点关联分析结果显示,AA基因型个体骨重显著大于AC基因型个体(P0.05)。除此之外,其余肉质性状指标及屠宰性状指标均差异不显著,暗示该SNPs位点可能对贵州白山羊骨重有影响。3.C-429T和A17110C的单体型分析表明,AACC、CACC和CACT单体型在提高系水力,CACT单体型在提高剪切力,AACC和CATT单体型在提高骨重,AACT、CACC和CACT单体型在提高肉骨比,AACC单体型在提高骨率方面可能发挥调控作用。4.肉质性能指标间相关分析表明,失水率与熟肉率和大理石纹均为极显著正相关(P0.01)。剪切力和系水力,大理石纹和系水力、熟肉率间呈显著正相关(P0.05)。屠宰性能指标间进行相关分析表明,活重与胴体重、净肉重、皮毛重、腰部肌肉厚极显著正相关(P0.01),与骨重显著正相关(P0.05),与皮率显著负相关(P0.05)。屠宰率与净肉重、腰部肌肉厚、净肉率、骨率、骨肉比极显著正相关(P0.01),与骨重显著相关(P0.05),与皮率极显著负相关(P0.05)。胴体重与屠宰率、净肉重、骨重、皮毛重、腰部肌肉厚、净肉率、骨肉比极显著正相关(P0.01),与骨率、骨肉比显著正相关(P0.05),与皮率极显著负相关(P0.01)。5.TFB2M基因生物信息学分析结果显示,A17110C位点处碱基变异致使其二级结构略微发生变化,最小自由能由-529.4 kJ/mol减少到-529.8 kJ/mol,结构稳定性略有增加。C-429T位于TFB2M基因的启动子区,预测发现存在2个可能的核心启动子区域。该SNPs位点使转录因子结合位点变化较大。同时,C-429T位点处于各软件预测的CpG岛区,但该位点对CPG岛的影响不大。6.实时荧光定量PCR检测TFAM基因和TFB2M基因在贵州白山羊各组织中的表达量。发现其表达规律分别是肺小脑垂体心下丘脑肾大脑肝脾;脾肾肺垂体大脑心小脑下丘脑肝。7.TFAM基因在肾组织、下丘脑、肺、大脑中的表达量分别与失水率、剪切力、胴体重、肉骨比呈显著正相关(P0.05),在垂体、肝脏中的表达量与系水力、骨重呈显著负相关(P0.05)。TFB2M基因在肺、脾、肾、垂体、肝脏、脾脏中的表达量与剪切力、失水率、肉色、大理石纹、大理石纹、大腿肌肉厚及活重呈显著正相关(P0.05),而在肝脏中的表达量与剪切力呈显著负相关(P0.05)。
[Abstract]:This study adopts PCR-SSCP to Guizhou white goat as the research object, DNA, direct sequencing, real-time fluorescence quantitative PCR, cloning method cDNA TFAM gene sequence and bioinformatics analysis of TFAM protein structure and physicochemical properties of the content, while exploring the TFAM and TFB2M gene mRNA expression and influence of polymorphism the Guizhou white goat meat traits and slaughter performance, for the development of Guizhou white goat meat, study of candidate genes related to slaughter performance, improving local goat meat quality, provide the basis for breeding. To speed up the test as the main achievements: cDNA sequence of TFAM gene in Guizhou white goats was 1. for the first time, through bioinformatics software analysis TFAM protein structure and physicochemical properties of.TFAM ORF protein 741 BP in length, encoding 246 amino acids. The theoretical isoelectric point of PI 9.74 showed that TFAM is acidic, at the same time, TFAM protein amino acid There is no sequence of transmembrane helix structure does not belong to the membrane protein or secretion of.TFAM protein analysis showed that TFAM protein was membrane function in cells most likely, value 13.18; gene ontology classification indicates that it may play a similar role for probabilistic numerical hormone 17.538. Guizhou white goat and sheep of the amino acid sequence homology of 98.8%.2. test screening of two SNPs loci of TFB2M gene were C-429T and A17110C respectively, two loci and three genotypes CC, CT and TT, AA, AC and CC. analysis showed that the genetic variation of two loci heterozygosity is high, polymorphism information content showed moderate polymorphism (0.25PIC0.50). 2 test show that the group, are consistent with the Hardy-Weinberg equilibrium of.TFAM gene to SNPs.C-429T no screening correlation analysis results showed that two loci, CC and CT genotype hydraulic system C-429T loci significantly Compared with the TT genotype (P0.05); the TT genotype was significantly greater than bone weight CT genotype (P0.05), suggesting that the SNPs site of Guizhou white goat meat and bone weight affect the results of.A17110C locus association analysis showed that AA genotype was significantly greater than bone weight AC genotype (P0.05). In addition, the meat quality traits and carcass traits index indexes were not significantly different, suggesting that the SNPs site of Guizhou white goat bone weight haplotype analysis showed that the effects of.3.C-429T and A17110C, AACC, CACC and CACT haplotypes in improving water stress, CACT haplotype in improving the shear stress, AACC and CATT haplotype in improving bone weight, AACT, CACC and CACT haplotype in improving meat bone ratio, AACC haplotype may play a regulatory role in.4. related performance analysis shows in improving bone meat rate, water loss rate and the rate of cooked meat and marbling were extremely significant 钁楁鐩稿叧(P0.01).鍓垏鍔涘拰绯绘按鍔，
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