發(fā)布時間：2018-05-18 12:10

  本文選題：荷斯坦奶牛 + 泌乳曲線��； 參考：《廣西大學》2015年碩士論文

【摘要】：本研究采用Wood等四種模型分別對荷斯坦奶牛泌乳曲線進行擬合,以期選出最佳擬合模型,為準確預測該牛場奶牛305d產(chǎn)奶量奠定基礎(chǔ),并為進一步研究荷斯坦奶牛的生產(chǎn)性能和泌乳規(guī)律提供參考依據(jù)；采用DNA池和PCR-SSCP法篩選AGPAT6和ABCA1基因SNP位點,結(jié)合HRM技術(shù)進行基因分型,研究基因變異和產(chǎn)奶量及奶成分的相關(guān)性；并進一步對AGPAT6基因mRNA的表達分析和啟動子甲基化分析,初步為揭示AGPAT6基因影響奶牛產(chǎn)奶性狀的分子調(diào)控機制提供理論基礎(chǔ)。研究結(jié)果如下：1.荷斯坦奶牛泌乳曲線的擬合分析利用SPSS17.0軟件對荷斯坦奶牛305d產(chǎn)奶量進行曲線擬合。結(jié)果表明：六次多項式模型為荷斯坦奶牛最佳泌乳曲線模型,其對各胎次泌乳曲線的擬合效果都較為理想。模型極顯著影響泌乳曲線的擬合度R2(P0.01),胎次對泌乳曲線擬合度R2無顯著性影響(P0.05)。2.AGPAT6基因多態(tài)性及其與產(chǎn)奶性狀的相關(guān)分析(1)AGPAT6基因8個(P1～P8)基因座中,在P1(exon1)、P2(exon3)、 P3 (intron3)、P6 (intron8)、P7 (intron9)和P8 (intron11)中各檢測到1個SNP,P4和P5中沒有檢測到多態(tài)性位點。每個SNP均有三種基因型。實驗牛群在這6個SNP s均處于中度多態(tài)(0.25PIC0.5)。(2)利用SPSS17.0軟件中的最小二乘法線性擬合模型對6個SNP s與奶牛產(chǎn)奶量和乳成分進行相關(guān)性分析。結(jié)果表明：P1位點不同基因型顯著影響乳成分(P0.05),P2、P3、P6和P8位點的不同基因型顯著影響產(chǎn)奶量和乳成分,P7位點不同基因型顯著影響產(chǎn)奶量、乳脂率和非脂固形物(P0.05),乳蛋白率無顯著差異(P0.05)。3.ABCA1基因多態(tài)性及其與產(chǎn)奶性狀的相關(guān)分析在ABCA1基因3個外顯子中,只在exon7中檢測到一個SNP。此位點有TT、TC和CC三種基因型。實驗牛群在此位點為中度多態(tài)(0.25PIC0.5),處于Hardy-Weinberg不平衡狀態(tài)。將此位點與荷斯坦奶牛的產(chǎn)奶性狀進行相關(guān)分析,發(fā)現(xiàn)不同基因型個體的產(chǎn)奶量存在極顯著差異(P0.01),而不同基因型個體乳脂率和乳蛋白率無顯著性差異(P0.05)。4.AGPAT6基因mRNA表達分析和啟動子甲基化分析(1)利用奶牛乳汁體細胞提取總RNA,研究同胎次產(chǎn)奶量接近而乳脂率不同的AGPAT6基因P2位點不同基因型個體mRNA表達水平。結(jié)果表明：TT型和TC型個體mRNA表達水平顯著高于CC型(P0.05),其mRNA表達水平與奶牛乳脂率正相關(guān)。(2)利用MS-PCR法分析AGPAT6基因P2位點不同基因型個體啟動子甲基化水平。結(jié)果表明：TT、TC和CC基因型個體啟動子甲基化狀態(tài)與基因表達未呈現(xiàn)一致的規(guī)律趨勢,表明P2位點不同基因型個體]mRNA表達水平可能不受AGPAT6基因啟動子甲基化過程的調(diào)控。
[Abstract]:In this study, Wood and other four models were used to fit the lactation curve of Holstein cows respectively, in order to select the best fitting model, and to lay the foundation for accurately predicting the milk yield of the dairy cows in 305 days. It also provides reference for further study on production performance and lactation law of Holstein cows, screening AGPAT6 and ABCA1 gene SNP loci by DNA pool and PCR-SSCP method, and carrying out genotyping with HRM technique. To study the correlation between gene variation and milk yield and milk composition, and to further analyze the expression of AGPAT6 gene mRNA and promoter methylation analysis, to provide a theoretical basis for revealing the molecular regulation mechanism of AGPAT6 gene affecting dairy cow milk production traits. The results are as follows: 1. Fitting Analysis of lactating Curve of Holstein Dairy cows the milk yield of Holstein cows at 305 days was fitted by SPSS17.0 software. The results showed that the six degree polynomial model was the best lactation curve model for Holstein cows, and the fitting effect of the curve was satisfactory. The model had a significant effect on the fitting degree of lactation curve R _ 2 / P _ (0.01), but the number of births had no significant effect on the fitness of milk curve R ~ (2). The polymorphism of AGPAT6 gene and its correlation with milk yield traits were analyzed in the locus of 8 (P1 / P8) gene of AGPAT6 gene, and the relationship between the polymorphism of AGPAT6 gene and lactation traits was analyzed. No polymorphic loci were detected in P1 exon1, P6 intron8, P7 intron9 and P8 intron11, respectively, in P1exon3, P3 intron3 and P6 intron8 intron9 and P8 intron11, respectively. No polymorphic loci were detected in P4 and P5. Each SNP has three genotypes. The correlation between six SNP s and milk yield and milk composition was analyzed by using the least square linear fitting model in SPSS17.0 software. The results showed that the different genotypes of different genotypes at the P1 locus significantly affected the milk yield and the milk yield of P0.05, P2, P3, P6 and P8 loci, and the milk yield was significantly affected by different genotypes of P7 locus. There was no significant difference between milk fat rate and non-fat solids (P0.05), and there was no significant difference in milk protein rate. The polymorphism of ABCA1 gene and its correlation with milk yield traits were detected only in exon7 and 3 exons of ABCA1 gene. There were three genotypes of TTTC and CC at this locus. The experimental cattle were in the Hardy-Weinberg imbalance state at this site, which was a moderate polymorphic 0.25PIC0.5. The correlation between this locus and milk yield traits of Holstein cows was analyzed. It was found that there were significant differences in milk yield among individuals with different genotypes (P 0.01), but there was no significant difference in milk fat and milk protein rates among individuals with different genotypes. AGPAT6 gene mRNA expression analysis and promoter methylation analysis were used to analyze milk somatic cells of dairy cows. Total RNAs were extracted to study the expression level of mRNA in individuals with different genotypes of AGPAT6 gene P2 locus with similar milk yield and different milk fat percentage. The results showed that the level of mRNA expression in individuals with different genotype P2 of AGPAT6 gene was significantly higher than that of P0.05 of CC type, and the mRNA expression level was positively correlated with milk fat rate of dairy cows.) the promoter methylation level of different genotype individuals with P2 locus of AGPAT6 gene was analyzed by MS-PCR method. The results showed that the methylation status of promoter in individuals with different genotypes of AGPAT6 and CC was not consistent with the trend of gene expression, indicating that the level of mRNA expression in individuals with different genotypes at P2 locus might not be regulated by the methylation process of promoter of AGPAT6 gene.
【學位授予單位】：廣西大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S823

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