本文關(guān)鍵詞： 貴州白山羊 肉質(zhì)性狀 屠宰性能 TFAM基因 TFB2M基因 關(guān)聯(lián)性分析　出處：《貴州大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：本研究以貴州白山羊?yàn)檠芯繉?duì)象,采用PCR-SSCP、DNA直接測(cè)序、實(shí)時(shí)熒光定量PCR、克隆等方法克隆得到TFAM基因的cDNA序列并利用生物信息學(xué)分析TFAM蛋白的結(jié)構(gòu)和理化性質(zhì)等內(nèi)容,同時(shí)探索了TFAM和TFB2M基因的mRNA表達(dá)規(guī)律和多態(tài)性對(duì)貴州白山羊肉質(zhì)性狀和屠宰性能的影響,為開展貴州白山羊肉質(zhì)、屠宰性能的候選基因相關(guān)研究,提高地方山羊羊肉品質(zhì),加快品種選育提供依據(jù)。試驗(yàn)獲得如下主要成果:1.首次得到的貴州白山羊TFAM基因的cDNA序列,通過生物信息學(xué)軟件分析TFAM蛋白的結(jié)構(gòu)和理化性質(zhì)。TFAM蛋白ORF長(zhǎng)741 bp,編碼246個(gè)氨基酸。其理論等電點(diǎn)PI為9.74表明TFAM是酸性的,同時(shí),TFAM蛋白氨基酸序列不存在跨膜螺旋結(jié)構(gòu)不屬于膜蛋白或分泌蛋白。TFAM蛋白功能分析表明TFAM蛋白在細(xì)胞被膜發(fā)揮功能的可能性最大,數(shù)值為13.18;基因本體分類表明,其可能發(fā)揮類似激素作用的概率數(shù)值為17.538。貴州白山羊和綿羊的氨基酸序列同源性為98.8%。2.試驗(yàn)篩選到TFB2M基因兩個(gè)SNPs位點(diǎn),分別是C-429T和A17110C,兩個(gè)位點(diǎn)分別出現(xiàn)三種基因型CC、CT和TT,AA、AC和CC。群體遺傳變異分析表明兩個(gè)位點(diǎn)雜合度均較高,多態(tài)信息含量均表現(xiàn)為中度多態(tài)(0.25PIC0.50)。χ2檢驗(yàn)表明,該群體在兩個(gè)位點(diǎn)均符合Hardy-Weinberg平衡狀態(tài)。TFAM基因沒有篩選到SNPs。C-429T關(guān)聯(lián)性分析結(jié)果顯示,C-429T位點(diǎn)的CC和CT基因型個(gè)體系水力顯著高于TT基因型個(gè)體(P0.05);TT基因型個(gè)體骨重顯著大于CT基因型個(gè)體(P0.05),暗示該SNPs位點(diǎn)可能對(duì)貴州白山羊的肉質(zhì)及骨重有影響。A17110C位點(diǎn)關(guān)聯(lián)分析結(jié)果顯示,AA基因型個(gè)體骨重顯著大于AC基因型個(gè)體(P0.05)。除此之外,其余肉質(zhì)性狀指標(biāo)及屠宰性狀指標(biāo)均差異不顯著,暗示該SNPs位點(diǎn)可能對(duì)貴州白山羊骨重有影響。3.C-429T和A17110C的單體型分析表明,AACC、CACC和CACT單體型在提高系水力,CACT單體型在提高剪切力,AACC和CATT單體型在提高骨重,AACT、CACC和CACT單體型在提高肉骨比,AACC單體型在提高骨率方面可能發(fā)揮調(diào)控作用。4.肉質(zhì)性能指標(biāo)間相關(guān)分析表明,失水率與熟肉率和大理石紋均為極顯著正相關(guān)(P0.01)。剪切力和系水力,大理石紋和系水力、熟肉率間呈顯著正相關(guān)(P0.05)。屠宰性能指標(biāo)間進(jìn)行相關(guān)分析表明,活重與胴體重、凈肉重、皮毛重、腰部肌肉厚極顯著正相關(guān)(P0.01),與骨重顯著正相關(guān)(P0.05),與皮率顯著負(fù)相關(guān)(P0.05)。屠宰率與凈肉重、腰部肌肉厚、凈肉率、骨率、骨肉比極顯著正相關(guān)(P0.01),與骨重顯著相關(guān)(P0.05),與皮率極顯著負(fù)相關(guān)(P0.05)。胴體重與屠宰率、凈肉重、骨重、皮毛重、腰部肌肉厚、凈肉率、骨肉比極顯著正相關(guān)(P0.01),與骨率、骨肉比顯著正相關(guān)(P0.05),與皮率極顯著負(fù)相關(guān)(P0.01)。5.TFB2M基因生物信息學(xué)分析結(jié)果顯示,A17110C位點(diǎn)處堿基變異致使其二級(jí)結(jié)構(gòu)略微發(fā)生變化,最小自由能由-529.4 kJ/mol減少到-529.8 kJ/mol,結(jié)構(gòu)穩(wěn)定性略有增加。C-429T位于TFB2M基因的啟動(dòng)子區(qū),預(yù)測(cè)發(fā)現(xiàn)存在2個(gè)可能的核心啟動(dòng)子區(qū)域。該SNPs位點(diǎn)使轉(zhuǎn)錄因子結(jié)合位點(diǎn)變化較大。同時(shí),C-429T位點(diǎn)處于各軟件預(yù)測(cè)的CpG島區(qū),但該位點(diǎn)對(duì)CPG島的影響不大。6.實(shí)時(shí)熒光定量PCR檢測(cè)TFAM基因和TFB2M基因在貴州白山羊各組織中的表達(dá)量。發(fā)現(xiàn)其表達(dá)規(guī)律分別是肺小腦垂體心下丘腦腎大腦肝脾;脾腎肺垂體大腦心小腦下丘腦肝。7.TFAM基因在腎組織、下丘腦、肺、大腦中的表達(dá)量分別與失水率、剪切力、胴體重、肉骨比呈顯著正相關(guān)(P0.05),在垂體、肝臟中的表達(dá)量與系水力、骨重呈顯著負(fù)相關(guān)(P0.05)。TFB2M基因在肺、脾、腎、垂體、肝臟、脾臟中的表達(dá)量與剪切力、失水率、肉色、大理石紋、大理石紋、大腿肌肉厚及活重呈顯著正相關(guān)(P0.05),而在肝臟中的表達(dá)量與剪切力呈顯著負(fù)相關(guān)(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垏鍔涘拰緋繪按鍔，

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