發(fā)布時間：2018-03-10 19:04

  本文選題：FASN　切入點(diǎn)：SCD1　出處：《西北農(nóng)林科技大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：脂肪酸合酶(Fatty acid synthase, FASN)是哺乳動物脂肪酸從頭合成的關(guān)鍵酶,催化短中鏈飽和脂肪酸的形成。硬脂酰輔酶A去飽和酶(Stearoyl-CoAdesaturase-1, SCD1)是催化形成單不飽和脂肪酸的關(guān)鍵酶,通過對飽和脂肪酸第9位碳鏈增加一個不飽和雙鍵而形成單不飽和脂肪酸,催化底物主要以棕櫚酸(C16：0)及硬脂酸(C18：0)為主,是乳腺自身進(jìn)行脂肪酸去飽和作用的關(guān)鍵酶。反芻動物乳脂的遺傳力較高,并且在乳腺組織中FASN和SCD1基因的mRNA表達(dá)量較高,因此本試驗(yàn)選取以上兩個基因作為候選基因。采集母羊分娩后4個月內(nèi)的奶樣(泌乳30天,60天,90天,120天),分析主要乳成分和脂肪酸組成,并與候選基因篩選出的多態(tài)位點(diǎn)進(jìn)行關(guān)聯(lián)分析。主要結(jié)果如下：1)通過DNA池測序技術(shù),分析得到FASN基因的3個多態(tài)位點(diǎn)(內(nèi)含子1的SNP C-911-T,內(nèi)含子2的SNP A-852-G,外顯子37的SNP T-14420-C)和SCD1基因的2個多態(tài)位點(diǎn)(外顯子3的SNP A-5205-G,內(nèi)含子3的SNP C-5311-T),并檢測到FASN基因存在兩個單倍型,H1 (C1-A2-T3)和H2 (T1-G2-C3)。所有突變位點(diǎn)中,除SNP A-5205-G外其他突變位點(diǎn)均符合哈代溫伯格平衡定律。利用線性混合模型,對FASN和SCD1基因的SNPs以及FASN基因的單倍型與產(chǎn)奶性狀進(jìn)行相關(guān)性分析。模型中涉及到種群效應(yīng)、檢測天數(shù)效應(yīng)、泌乳時間效應(yīng)、胎次效應(yīng)、個體效應(yīng)、公畜效應(yīng)、等位基因效應(yīng)和單倍型效應(yīng)以及泌乳時間內(nèi)等位基因與單倍型之間的互作效應(yīng)。FASN基因的3個SNP在不同泌乳時間均顯著影響了C6：0,C8：0,C10：0和C14：0脂肪酸的含量,并且主要等位基因均顯著下調(diào)了從頭合成的脂肪酸含量。單倍型H1(C1A2T3)也顯著下調(diào)了從頭合成的脂肪酸含量,但是單倍型H2(T1G2C3)對從頭合成脂肪酸的影響與之相反。此外,單倍型H2顯著性影響了C18:1 n9 trans和CLA cis9, tran11的含量。SCD1基因的SNP位點(diǎn)C-5311-T在不同泌乳時間均顯著影響了C16：1,C13：0,C14：1脂肪酸的含量。盡管SCD1基因的SNP A-5205-G不符合哈代溫伯格平衡定量,但是其對C4：0,C11：0,C12：0,C16：1,C17：1,C18：0脂肪酸的含量具有顯著性影響。以上結(jié)果表明,這些多態(tài)位點(diǎn)可以作為改善山羊奶品質(zhì)的分子標(biāo)記。2)利用線性混合模型分析泌乳時間、胎次、種群采樣時間等因素對羊奶成分以及脂肪酸組成的影響。模型分析過程中涉及的隨機(jī)效應(yīng)包括公畜和羊只個體的效應(yīng)。采樣時間和種群因素顯著影響乳中脂肪酸的組成,表明飼養(yǎng)管理水平和環(huán)境因素對乳成分以及乳中脂肪酸組成具有重要的影響。隨著泌乳時間的延長以及胎次的增加,羊奶中飽和脂肪酸含量增加,與其相反,單不飽和脂肪酸和多不飽和脂肪酸含量降低。結(jié)果表明,山羊乳腺組織的發(fā)育隨著年齡的增長趨向完善。隨著泌乳時間的延長,脂肪與蛋白的校正比值出現(xiàn)顯著下調(diào)。表明在奶山羊泌乳早期脂解作用的存在。在早期泌乳階段,奶中多不飽和脂肪酸含量更為豐富,而飽和脂肪酸含量較少。隨著泌乳時間的延長,CLA cis9,trins11的含量增加,推測隨著泌乳時間的延長,SCD1的活性及表達(dá)水平不斷升高。相關(guān)性分析表明,從頭合成的飽和脂肪酸與C18類多不飽和脂肪酸存在負(fù)相關(guān),這可能是由于多不飽和脂肪酸對脂肪酸從頭合成的抑制作用所致。盡管C4：0脂肪酸被認(rèn)定是從頭合成而來,但是其與中鏈脂肪酸存在負(fù)相關(guān),說明C4：0脂肪酸的來源不止一條途徑。奶中的奇數(shù)鏈脂肪酸主要來源于瘤胃中的微生物,因此其含量以及與其他脂肪酸的相關(guān)性反應(yīng)了瘤胃微生物群落的活性。奇數(shù)鏈脂肪酸與CLA cis9, trans 11以及其他反式脂肪酸存在正相關(guān),說明奇數(shù)鏈脂肪酸與反式脂肪酸的相關(guān)性可以作為診斷奶山羊瘤胃微生物活性的一種生物學(xué)標(biāo)記。本試驗(yàn)首次分析了影響奶中脂肪酸變化的因素,為日后改善羊奶品質(zhì)提供了理論依據(jù)。
[Abstract]:Fatty acid synthase (Fatty acid, synthase, FASN) is a key enzyme in mammalian fatty acid biosynthesis, the formation of short chain saturated fatty acid catalysis. Stearoyl coenzyme A desaturase (Stearoyl-CoAdesaturase-1, SCD1) is the key enzyme catalyzing the formation of monounsaturated fatty acids and saturated fatty acids by ninth carbon chain add an unsaturated double bond and the formation of monounsaturated fatty acids catalyzed mainly by palmitic acid (C16:0) and stearic acid (C18:0), is a key enzyme in breast self fatty acid desaturation. Fat of ruminant animal genetic capacity is high, and FASN in breast tissue and SCD1 gene mRNA the expression level is high, so this experiment selected above two gene as candidate gene. The milk samples within 4 months after birth (lactation ewes collected 30 days, 60 days, 90 days, 120 days), the main composition analysis of milk composition and fatty acid, and the candidate base Polymorphic loci were screened by correlation analysis. The main results are as follows: 1) by DNA sequencing technology, analysis of 3 polymorphisms of FASN gene (SNP, C-911-T, SNP in intron 1 A-852-G and intron 2 and exon 37 of SNP T-14420-C) 2 polymorphic sites and SCD1 gene (the exon 3 of SNP A-5205-G, SNP C-5311-T), and intron 3 of FASN gene was detected in two haplotypes, H1 (C1-A2-T3) and H2 (T1-G2-C3). All mutations, in addition to SNP A-5205-G the other mutations are in line with the Hardy Weinberg equilibrium. Using linear mixed model, correlation analysis the SNPs of FASN and SCD1 gene and FASN gene haplotypes and milk production traits. Related to the population effect model, testing days effect, lactation time effect, birth order effect, individual effect, sire effect, allele effect and haplotypic effect and time of lactation In between the allele and haplotype interaction of 3 SNP effect of.FASN gene in different lactation time had significant effects on C6:0, C8:0, C10:0 and C14:0 fatty acid content, and the main allele were significantly reduced the fatty acid content of de novo synthesized. Haplotype H1 (C1A2T3) also significantly lowered the fatty acid the content of de novo synthesized, but the haplotype of H2 (T1G2C3) on the de novo synthesis of fatty acids on the contrary. In addition, the haplotype of H2 significantly affected C18:1 N9 trans and CLA cis9, the content of.SCD1 tran11 gene SNP locus C-5311-T in different lactation time were significantly affected by C16:1, C13:0, the content of fatty acid C14:1 although the SCD1 gene SNP. A-5205-G does not comply with the Hardy Weinberg equilibrium quantity, but the C4:0, C11:0, C12:0, C16:1, C17:1, significantly influenced the fatty acid content of C18:0. The above results suggest that this Some polymorphic loci can be used as a molecular marker of.2 improve the goat milk quality) of lactation time, using linear mixed model parity, sampling time and other factors on the impact of population goats'milk composition and fatty acid composition. The random effects model analysis involving process including the effect of sire and sheep. The individual sampling time and significantly influence the composition of population the fatty acids in milk, feeding management and environmental factors that have important influence on milk composition and fatty acid composition in milk. With prolonged lactation time and increased parity, increase in saturated fatty acid content of goats' milk instead, monounsaturated fatty acid and polyunsaturated fatty acid content decreased. Show that the goat mammary tissue development along with the age growth tendency to improve. With prolonged lactation time, ratio of fat and protein correction was reduced. In that Early lactation dairy goat lipolysis exists. In the early stage of lactation, milk polyunsaturated fatty acid content is more abundant, and the saturated fatty acid content is low. With prolonged lactation time, CLA cis9, trins11 content increased, presumably with prolonged lactation time, the activity and the expression of SCD1 increased. The correlation analysis shows that the de novo synthesis of acid saturated fat and C18 polyunsaturated fatty acids are negatively correlated, this may be due to polyunsaturated fatty acids on inhibition of fatty acid induced de novo synthesis of fatty acids were identified. Although C4:0 is from the head to the synthesis, but with medium chain fatty acids are negatively correlated. Explain the source of C4:0 fatty acids in more than one way. The odd chain fatty acids in milk from the main source of rumen microorganisms, so the correlation between the contents and fatty acid reaction of rumen microbial communities The activity of odd chain fatty acid and CLA cis9, trans 11 and other trans fatty acids were positively correlated, indicating the correlation of odd chain fatty acids and trans fatty acids can be used as a biomarker for the diagnosis of dairy goat rumen microbial activity. This is the first analysis of factors influencing changes in milk fatty acids, provide the theoretical basis for improving the quality of goats'milk day.

【學(xué)位授予單位】：西北農(nóng)林科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S827

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