本文選題：牦牛 + KRTAP6家族基因　； 參考：《甘肅農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：角蛋白關(guān)聯(lián)蛋白(Keratins-associated proteins,KRTAPs)是毛纖維的主要結(jié)構(gòu)成分,由KRTAPs基因家族編碼,KARTAP6基因家族屬于高甘氨酸/酪氨酸角蛋白(HGT-KRTAPs)基因家族,其突變影響毛纖維細(xì)度。脂肪酸合成酶(Fatty acid synthase,FASN)是脂肪酸合成代謝的關(guān)鍵酶,在動物體脂和乳脂的脂肪酸合成過程中發(fā)揮著關(guān)鍵作用。本研究應(yīng)用PCR-SSCP技術(shù)檢測甘南牦牛、天祝白牦牛、青海牦牛和野血牦牛KRTAP6家族基因編碼區(qū)及FASN基因啟動子區(qū)、第34外顯子突變,分析其多態(tài)性和序列變異特征;結(jié)合甘南牦牛乳品質(zhì)性狀測定,分析FASN基因突變對乳品質(zhì)性狀的影響。主要結(jié)果為:1.牦牛KRTAP6家族基因編碼區(qū)多態(tài)性較豐富。(1)KRTAP6-1基因編碼區(qū)檢測到A_1、B_1、C_1、D_1、E_1 5種等位基因,序列比對發(fā)現(xiàn)4處SNPs及1處45bp的插入/缺失突變,其中非同義突變c.173GC和c.175GT導(dǎo)致氨基酸殘基p.Cys58Ser和p.Gly59Cys的改變,45bp的插入/缺失突變導(dǎo)致16個氨基酸殘基的缺失;等位基因A_1頻率56.5%~76.0%,為優(yōu)勢等位基因;甘南牦牛PIC值0.222呈低度多態(tài),其他3個牦牛群體PIC值0.393~0.467呈中度多態(tài)。(2)牦牛KRTAP6-2基因編碼區(qū)檢測到A_2、B_2和C_23種等位基因,序列比對發(fā)現(xiàn)3處SNPs和1處6bp的插入/缺失,其中非同義突變c.248AG導(dǎo)致p.Tyr19Cys的氨基酸殘基改變,6bp的插入/缺失突變導(dǎo)致2個氨基酸殘基的缺失;等位基因A_2頻率73.2%~87.3%,為優(yōu)勢等位基因;甘南牦牛PIC值0.387呈中度多態(tài),其他3個牦牛群體PIC值0.203~0.246呈低度多態(tài)。(3)牦牛KRTAP6-4基因編碼區(qū)檢測到A_3、B_3、C_3、D_3和E_3 5種等位基因,序列比對發(fā)現(xiàn)6處SNPs,其中c.118AG的非同義突變導(dǎo)致編碼氨基酸殘基p.Ser40Gly的改變;等位基因A_3頻率73.7%~82.8%為優(yōu)勢等位基因;4個牦牛群體PIC值0.265~0.387呈中度多態(tài)。(4)甘南牦牛KRTAP6-1、KRTAP6-2和KRTAP6-4基因編碼區(qū)顯著偏離Hardy-Weinberg平衡狀態(tài)。2.序列比對及聚類分析發(fā)現(xiàn)牦牛KRTAP6-1、KRTAP6-2和KRTAP6-4基因位于第1號染色體上;牦牛KRTAP6-1基因外顯子區(qū)堿基序列與普通牛、山羊和綿羊的同源性最高,系統(tǒng)進(jìn)化與其親緣關(guān)系遠(yuǎn)近一致。3.牦牛FSAN基因啟動子區(qū)和第34外顯子多態(tài)性較豐富。(1)啟動子區(qū)檢測到3種等位基因A,B和C,序列比對發(fā)現(xiàn)c.1-1948.TC和c.1-1910CA的堿基突變;等位基因A頻率74.4%~75.2%為優(yōu)勢等位基因;PIC值0.366~0.376為中度多態(tài)。(2)第34外顯子區(qū)檢測到2種等位基因D和E,序列比對發(fā)現(xiàn)c.15281+74 AT的堿基突變;等位基因D頻率77.6%~79.5%為優(yōu)勢等位基因;3個牦牛群體PIC值0.272~0.286為中度多態(tài)。(3)啟動子區(qū)和第34外顯子區(qū)突變位點(diǎn)間形成4種單倍型,其中單倍型D1A2頻率最高為66.74%,D1B2型頻率最低為6.96%;兩區(qū)域突變位點(diǎn)間存在一定程度連鎖關(guān)系且接近連鎖平衡狀態(tài)。4.FASN基因突變影響甘南牦牛部分乳品質(zhì)性狀。啟動子區(qū)基因型AA個體的乳脂率顯著高于基因型AB和AC個體(P0.05);第34外顯子區(qū)基因型DD個體乳脂率顯著高于基因型DE個體(P0.05),而基因型DE個體乳糖百分含量顯著高于DD型個體(P0.05)。
[Abstract]:Keratins-associated proteins (KRTAPs) is the main structural component of wool fiber, which is encoded by the KRTAPs gene family. The KARTAP6 gene family belongs to the high glycine / tyrosine keratin (HGT-KRTAPs) gene family, and its mutation affects the fineness of the wool fiber. The fatty acid synthase (Fatty acid synthase, FASN) is a fatty acid synthesis generation. The key enzyme of Xie is playing a key role in the synthesis of fatty acid in animal body fat and milk fat. In this study, the PCR-SSCP technique was used to detect the gene coding region of Gannan yak, Tianzhu white yak, Qinghai yak and wild blood yak, and FASN gene promoter region, thirty-fourth exon mutation, analysis of polymorphism and sequence variation characteristics; The effects of FASN gene mutation on the quality of milk quality were analyzed in Gannan yak milk quality traits. The main results were: 1. yak KRTAP6 family gene coding region polymorphism was rich. (1) KRTAP6-1 gene coding region detected 5 alleles A_1, B_1, C_1, D_1, E_1, sequence alignment found 4 SNPs and 1 45bp insertion / deletion mutation, which is not The synonymous mutations c.173GC and c.175GT lead to changes in the amino acid residues p.Cys58Ser and p.Gly59Cys, and the insertion / deletion mutation of 45bp leads to the deletion of the 16 amino acid residues; the A_1 frequency of the allele 56.5%~76.0% is the dominant allele; the PIC value of the Gannan yak is 0.222 low polymorphism, and the PIC value 0.393~0.467 of the other 3 yak populations is moderately polymorphic. (2) A_2, B_2 and C_23 alleles were detected in the yak KRTAP6-2 gene coding region. Sequence alignment found 3 SNPs and 1 6BP insertion / deletion, in which the non synonymous mutation c.248AG leads to the change of the amino acid residues in p.Tyr19Cys, and the insertion / deletion mutation of 6BP leads to the loss of 2 amino acid residues, and the allele A_2 frequency 73.2%~87.3% is the dominant allele. The PIC value 0.387 of Gannan yak was moderately polymorphic and the PIC value 0.203~0.246 of the other 3 yak populations showed low polymorphism. (3) the 5 alleles of A_3, B_3, C_3, D_3 and E_3 were detected in the yak KRTAP6-4 gene coding region, and the sequence alignment found 6 SNPs, in which the non synonymous process of c.118AG resulted in the alteration of the encoded amino acid residues; The _3 frequency 73.7%~82.8% was the dominant allele; the PIC value of 4 yak populations was moderately polymorphic. (4) KRTAP6-1, KRTAP6-2 and KRTAP6-4 genes in Gannan yaks, KRTAP6-2 and KRTAP6-4 genes were significantly deviated from the.2. sequence alignment of Hardy-Weinberg and cluster analysis found that the Yak's KRTAP6-1, KRTAP6-2 and KRTAP6-4 genes were on the first chromosome; The nucleotide sequence of the exons of the 6-1 gene was the highest with the common cattle, goats and sheep. The phylogenetic relationship was far and close to the.3. yak FSAN gene promoter and thirty-fourth exon polymorphism. (1) the promoter region detected 3 alleles A, B and C, and the sequence alignment found the base mutation of c.1-1948.TC and c.1-1910CA; The allele A frequency 74.4%~75.2% was the dominant allele; PIC 0.366~0.376 was moderate polymorphism. (2) 2 alleles D and E were detected in thirty-fourth exons, sequence alignment found the base mutation of c.15281+74 AT; the D frequency 77.6%~79.5% of the allele was the dominant allele; 3 yak populations PIC 0.272~0.286 was moderate polymorphism. (3) promoter region and (3) promoter region. Thirty-fourth exon mutation sites formed 4 haplotypes, in which the frequency of haplotype D1A2 was the highest and the D1B2 type frequency was the lowest 6.96%; there was a certain degree of linkage between the two loci and the linkage equilibrium state.4.FASN gene mutation affected the milk quality traits of Gannan yak. The milk fat of the promoter region genotype AA individual The rate was significantly higher than that of genotype AB and AC individuals (P0.05); thirty-fourth exon genotype DD individual milk fat rate was significantly higher than that of genotype DE individuals (P0.05), while the content of lactose in genotype DE was significantly higher than that of DD type individuals (P0.05).
【學(xué)位授予單位】：甘肅農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S823.85

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