本文關(guān)鍵詞： 奶牛 HSP70-A8 遺傳多態(tài)性 表達水平 DHI性能 生理指標　出處：《安徽農(nóng)業(yè)大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：熱應(yīng)激蛋白70(HSP70)作為最重要的分子伴侶參與到動物機體內(nèi)的熱應(yīng)激反應(yīng)之中,為了研究HSP70-A8基因?qū)δ膛Ｉa(chǎn)性能和耐熱性能的影響,以397頭荷斯坦奶牛為主要研究對象,采用單鏈構(gòu)象多態(tài)性(PCR-SSCP)技術(shù),對HSP70-A8基因編碼區(qū)(664 to 5089)進行克隆與多態(tài)性研究,并分析其與奶牛DHI生產(chǎn)性能的相關(guān)性；以10頭荷斯坦牛和娟×荷雜交牛為研究對象,采用實時熒光定量技術(shù)(RT-PCR)分析和比較了不同遺傳型奶牛在不同季節(jié)(環(huán)境溫度)下HSP70-A8基因在血液組織中的相對表達水平,同時分析了表達水平與生理指標的關(guān)系。取得的結(jié)果如下：1.荷斯坦奶牛HSP70-A8基因的多態(tài)性及其遺傳分析利用PCR-SSCP方法進行檢測,在HSP70-A8基因的第2-8外顯子上均沒有檢測到突變,在第1外顯子1522BP處存在G→C的錯義突變,使編碼的氨基酸由谷氨酰胺變成組氨酸。檢測到有三種基因型AA、AB和BB。其中AA基因型頻率為38.56%,AB基因型頻率分別為31.03%,BB基因型頻率分別為30.41%；多態(tài)信息含量(PIC)為0.3733,在該位點均處于中度多態(tài)；卡方檢驗表明試驗牛群體處于哈迪-溫伯格不平衡狀態(tài)。2.荷斯坦奶牛HSP70-A8基因的多態(tài)性與生產(chǎn)性能的關(guān)聯(lián)分析對該位點的不同基因型與生產(chǎn)性能進行關(guān)聯(lián)分析,結(jié)果表明：AA型和AB型個體的305d標準乳產(chǎn)量、總經(jīng)濟校正奶量、305d乳脂產(chǎn)量、305d乳蛋白產(chǎn)量和總?cè)榈鞍桩a(chǎn)量極顯著低于BB型個體(P0.01)；AA型個體的標準乳總產(chǎn)量、總產(chǎn)奶量、305d經(jīng)濟校正奶量和總?cè)橹a(chǎn)量顯著低于AB型個體(P0.05),AB型又極顯著低于BB型個體(JP0.01);AA型個體的體細胞數(shù)極顯著低于AB型個體和BB型個體(P0.01)。3.不同遺傳型奶牛在不同環(huán)境溫度下HSP70-A8基因表達水平的比較分析采用實時熒光定量PCR方法分析HSP70-A8基因在不同季節(jié)中的相對表達量,結(jié)果表明：夏季HSP70-A8基因在荷斯坦牛和F1代中的表達量極顯著低于在F2代中的表達量(P0.01),秋季和冬季HSP70-A8基因在三個品種中的表達差異不顯著；荷斯坦奶牛HSP70-A8基因在三個季節(jié)中的表達量差異不顯著,F1代HSP70-A8基因在三個季節(jié)的表達量差異不顯著,F2代HSP70-A8基因在夏季的表達量顯著高于冬季(P0.01)。4.奶牛主要生理指標差異性分析及其與HSP70-A8基因表達水平關(guān)聯(lián)分析對HSP70-A8基因表達量和生理指標進行分析,結(jié)果表明：奶牛在夏季的呼吸頻率顯著高于秋冬季節(jié)(P0.01),夏季的直腸溫度明顯高于冬季(P0.01),奶牛在三個季節(jié)中的心跳次數(shù)差異不顯著。在三個季節(jié)中,不同品種之間的呼吸頻率、直腸溫度和心跳次數(shù)差異均不顯著。HSP70-A8的表達量和奶牛的呼吸頻率、心跳次數(shù)和直腸溫度均無明顯相關(guān)性。研究結(jié)果提示HSP70-A8基因?qū)τ谀膛５纳a(chǎn)性能具有顯著性影響,可以作為奶牛生產(chǎn)性能選擇的候選遺傳分子標記,BB基因型個體可以作為生產(chǎn)性狀優(yōu)勢個體進行培育。
[Abstract]:Heat stress protein 70 (HSP70), as the most important molecular chaperone, is involved in the heat stress reaction in animal body. In order to study the effect of HSP70-A8 gene on the performance and heat resistance of dairy cattle, single strand conformation polymorphism (SSCP) technique was used to study 397 Holstein cows. Cloning and polymorphism of HSP70-A8 gene coding region from 664 to 5089) were studied, and the correlation between DHI production performance and DHI was analyzed. Ten Holstein cattle and Juan 脳 Hollow hybrid cattle were studied. The relative expression level of HSP70-A8 gene in blood tissues of dairy cattle with different genetic types in different seasons (ambient temperature) was analyzed and compared by real-time fluorescence quantitative technique RT-PCR. At the same time, the relationship between expression level and physiological indexes was analyzed. The results were as follows: 1. The polymorphism of HSP70-A8 gene in Holstein cows and its genetic analysis were detected by PCR-SSCP method. No mutation was detected in exon 2-8 of HSP70-A8 gene, and G existed at exon 1522BP of exon 1. 鈫扵he missense mutation of C resulted in the transformation of the encoded amino acid from glutamine to histidine. Three genotypes were detected, AAAAB and BB.The frequency of AA genotype was 38.56%. The frequency of AB genotype was 31.03and BB genotype was 30.41; The polymorphic information content (Pi) was 0.3733, which was moderately polymorphic at this locus. The chi-square test showed that the population of the tested cattle was in the Hady-Weinberg disequilibrium state. 2. Correlation analysis of the polymorphism of HSP70-A8 gene and the production performance of Holstein dairy cattle with different genotypes and productivity of the locus. To be able to perform correlation analysis. The results showed that the 305d standard milk yield and the total corrected milk yield were 305d and 305d respectively. The milk protein yield and total milk protein yield at 305d were significantly lower than those of BB type individuals (P 0.01). The total standard milk yield, total milk yield and total milk fat yield of AA individuals were significantly lower than those of AB individuals (P 0.05). AB type was significantly lower than BB type individuals. The number of somatic cells in AA individuals was significantly lower than that in AB individuals and BB individuals (P 0.01). Comparative analysis of HSP70-A8 gene expression levels in different genotypes of dairy cattle under different ambient temperature using real-time fluorescence quantitative PCR method to analyze the phase of HSP70-A8 gene in different seasons. Yes, expression. The results showed that the expression of HSP70-A8 gene in Holstein cattle and F1 generation in summer was significantly lower than that in F2 generation (P 0.01). There was no significant difference in the expression of HSP70-A8 gene between autumn and winter. There was no significant difference in the expression of HSP70-A8 gene among the three seasons in Holstein dairy cattle. There was no significant difference in the expression of HSP70-A8 gene in F1 generation in the three seasons. The expression of HSP70-A8 gene in F2 generation in summer was significantly higher than that in winter (P0.01). Analysis of the difference of the main physiological indexes and the correlation analysis between the expression level of HSP70-A8 gene and the expression level of HSP70-A8 gene in dairy cattle. The expression quantity and physiological index of HSP70-A8 gene were analyzed. The results showed that the respiratory frequency in summer was significantly higher than that in autumn and winter, and the rectal temperature in summer was significantly higher than that in winter. There was no significant difference in the number of heart beats in the three seasons. In the three seasons, the respiration rate among different breeds was not significant. There was no significant difference in rectal temperature and heart rate. The expression of HSP70-A8 and the respiratory rate of dairy cattle were not significant. There was no significant correlation between heart rate and rectal temperature. The results showed that HSP70-A8 gene had a significant effect on the performance of dairy cattle. The individuals of BB genotype which can be used as candidate genetic markers for performance selection of dairy cattle can be bred as dominant individuals of production traits.
【學(xué)位授予單位】：安徽農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S823

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