發(fā)布時(shí)間：2018-04-18 14:13

  本文選題：家馬 + 產(chǎn)奶量�。� 參考：《西北農(nóng)林科技大學(xué)》2017年碩士論文

【摘要】：馬奶含有豐富的維生素和礦物質(zhì),營(yíng)養(yǎng)價(jià)值極高,但其產(chǎn)量遠(yuǎn)低于牛奶。隨著人民生活水平的提高,馬奶的需求量增加,加強(qiáng)馬泌乳遺傳基礎(chǔ)研究,開發(fā)相關(guān)的分子標(biāo)記,培育乳用馬品種勢(shì)在必行。本研究選擇了9個(gè)家畜泌乳性能候選基因:催乳素(PRL)及催乳素受體基因(PRLR)、垂體特異性轉(zhuǎn)錄因子1(POU1F1)、生長(zhǎng)激素1(GH1)及生長(zhǎng)激素受體基因(GHR)、乳鐵蛋白基因(LTF)、雌激素受體基因(ESR1、ESR2)和三磷酸腺苷結(jié)合轉(zhuǎn)運(yùn)蛋白G超家族成員2基因(ABCG2),根據(jù)NCBI提供的基因序列設(shè)計(jì)擴(kuò)增CDS區(qū)序列,以3個(gè)中國(guó)家馬品種(伊犁馬、關(guān)中馬和蒙古馬)共337匹馬為研究對(duì)象,測(cè)序檢測(cè)鑒定候選基因CDS區(qū)的單核苷酸多態(tài)位點(diǎn),采用PCR-RFLP技術(shù)對(duì)其進(jìn)行分型,并將伊犁馬上發(fā)現(xiàn)SNP位點(diǎn)與伊犁馬日均產(chǎn)奶量進(jìn)行單因素方差分析,獲得如下研究結(jié)果:1.在3個(gè)中國(guó)家馬品種9個(gè)泌乳相關(guān)基因中檢測(cè)到10個(gè)SNPs(PRL c.210 CT,PRL g.9115 TG,PRL g.9119 CA,PRL g.11090 AG,PRLR g.145227 TG,POU1F1g.2758 TC,GH1 c.22 AC,GH1 g.1584 CG,GHR g.-562 CT,LTF c.172 GA),均為首次發(fā)現(xiàn)。其中PRLR g.145227 TG,GHR g.-562 CT,LTF c.172 GA 3個(gè)位點(diǎn)僅發(fā)現(xiàn)于伊犁馬上,PRL g.9115 TG,PRL g.11090 AG,GH1 c.22 AC,GH1 g.1584 CG4個(gè)位點(diǎn)僅發(fā)現(xiàn)于關(guān)中馬上,PRL g.9119 CA位點(diǎn)為關(guān)中馬、蒙古馬共有,PRL c.210CT,POU1F1 g.2758 TC位點(diǎn)在三個(gè)品種上均有發(fā)現(xiàn)。全部10個(gè)SNP多態(tài)位點(diǎn)中,有3個(gè)位點(diǎn)發(fā)生在基因的CDS區(qū)域,引起錯(cuò)義突變;1個(gè)位點(diǎn)發(fā)生在基因的啟動(dòng)子區(qū)域,其余6個(gè)位點(diǎn)均發(fā)生在內(nèi)含子上。2.使用PCR-RFLP方法對(duì)伊犁馬上檢測(cè)到的SNP多態(tài)進(jìn)行分型統(tǒng)計(jì),并與其120天日均產(chǎn)奶量進(jìn)行關(guān)聯(lián)分析。檢測(cè)到伊犁馬上存在的5個(gè)不同基因的SNP位點(diǎn)中,PRL c.210 CT和LTF c.172 GA是發(fā)生在CDS區(qū)上的錯(cuò)義突變,PRL c.210 CT突變導(dǎo)致PRL基因編碼蛋白第70位苯丙氨酸突變?yōu)榱涟彼?LTF c.172 GA突變致LTF編碼蛋白第57位精氨酸突變?yōu)橘嚢彼?GHR g.-562 CT發(fā)生在GHR基因啟動(dòng)子區(qū)域,其余位點(diǎn)均位于內(nèi)含子上。與日均產(chǎn)奶量的關(guān)聯(lián)分析結(jié)果顯示,5個(gè)不同基因的SNP位點(diǎn)中,除PRL c.210 CT外,各位點(diǎn)不同基因型的日均產(chǎn)奶量差異不顯著。PRL c.210 CT位點(diǎn)不同基因型的日均產(chǎn)奶量差異顯著,其中CC基因型個(gè)體日均產(chǎn)奶量顯著高于CT基因型個(gè)體,CT基因型個(gè)體日均產(chǎn)奶量與TT基因型個(gè)體差異不顯著,推測(cè)C等位基因可能對(duì)伊犁馬產(chǎn)奶量性狀有正遺傳效應(yīng)。3.使用雙熒光素酶報(bào)告基因系統(tǒng),檢測(cè)馬PRL、PRLR基因啟動(dòng)子活性區(qū)域。檢測(cè)結(jié)果顯示,馬PRL基因上游-418~-1050bp區(qū)間為該基因的主要活性區(qū)域,其中-418~-738bp區(qū)間存在較強(qiáng)的順勢(shì)作用元件;馬PRLR基因啟動(dòng)子活性區(qū)域位于上游-702~-2399bp區(qū)間,-1546~-1798bp區(qū)間存在較強(qiáng)的順勢(shì)作用元件。目前,中國(guó)家馬品種泌乳性能相關(guān)基因的研究甚少。本研究初步調(diào)查了3個(gè)中國(guó)家馬品種9個(gè)泌乳相關(guān)基因CDS區(qū)SNP情況,并將伊犁馬上發(fā)現(xiàn)SNP位點(diǎn)與其日均產(chǎn)奶量進(jìn)行關(guān)聯(lián)分析,初步探索了馬PRL、PRLR基因啟動(dòng)子活性區(qū)域,為中國(guó)乳用馬的高效選育、種質(zhì)資源保存與利用提供了一定的依據(jù)。
[Abstract]:Milk is rich in vitamins and minerals, high nutritional value, but its yield is much lower than that of milk. With the improvement of people's living standard, demand for milk increase, to strengthen the study of the genetic basis of horse lactation, development of molecular markers related to the breeding of dairy horse breeds is imperative. This study selected 9 livestock lactation performance candidate genes: prolactin (PRL) and prolactin receptor gene (PRLR), pituitary specific transcription factor 1 (POU1F1) 1 (GH1), growth hormone and growth hormone receptor gene (GHR), lactoferrin (LTF) gene, estrogen receptor gene (ESR1, ESR2) and ATP binding cassette superfamily G members of the 2 gene (ABCG2), according to the amplification of sequences of CDS gene sequence design provided by NCBI, with 3 China home horse breeds (Yili horse, Mongolia horse and horse Guanzhong) a total of 337 horses as the research object, the detection and identification of candidate gene CDS region nucleotide sequencing Polymorphic loci, carries on the classification using PCR-RFLP technology, and Yili will immediately find SNP site and Yili Marie are milk production by single factor variance analysis, the main results are as follows: 10 to 1. in 9 lactation SNPs detected 3 genes related to Chinese horse breeds (PRL c.210 CT, PRL g.9115 TG PRL, g.9119 CA, PRL g.11090 AG, PRLR g.145227 TG, POU1F1g.2758 TC, GH1 c.22 AC, GH1 g.1584 CG, GHR g.-562 CT, LTF c.172, GA) were found for the first time. The PRLR g.145227 TG, GHR g.-562 CT, LTF c.172 GA 3 loci was found immediately PRL g.9115 TG Li huy, PRL. G.11090 AG, GH1 c.22 AC, GH1 g.1584 CG4 loci were found only in the PRL g.9119 CA site immediately, for a total of Guanzhong horse, Mongolia horse, PRL c.210CT, POU1F1 g.2758 TC loci were found in three cultivars. All 10 SNP loci, 3 loci in CDS gene Area caused by missense mutations; 1 loci in the promoter regions of genes, the remaining 6 loci occurred in the intron of.2. using the PCR-RFLP method to detect the SNP polymorphism in Yili immediately classified statistics, and the 120 day average milk production for the correlation analysis. SNP loci of 5 different genes detected Yili immediately exists in the PRL c.210 CT and LTF c.172 GA in CDS region is a missense mutation, PRL c.210 CT mutation in the PRL gene encoding protein seventieth mutated to phenylalanine leucine, LTF c.172 GA induced LTF mutation protein encoding fifty-seventh mutation of arginine to lysine; GHR g.-562 CT start the sub region in the GHR gene, the other sites were located in introns. The correlation analysis result and average daily milk yield showed that 5 different gene loci of SNP, c.210 in addition to PRL CT, average daily milk yield differences between different genotypes have not The average daily milk yield were.PRL c.210 CT loci had significant difference, the CC genotype average daily milk yield was significantly higher than that of the CT genotype, CT genotype average daily milk yield and TT genotypes were not significantly different, suggesting that C allele may have positive genetic effect of.3. using dual luciferase gene system on Yili horse milk production traits, detection of equine PRL, PRLR gene promoter region. Test results show that the horse PRL gene upstream -418~-1050bp interval as the main active region of the -418~-738bp gene, which has a strong range of cis acting element; Ma PRLR gene promoter region located upstream of the -702~-2399bp interval, -1546~-1798bp interval exists homeopathic components strong. At present, the research Chinese home lactation performance of horse breeds related genes understood. We investigated the 3 Chinese horse breed 9 mi The SNP status in the CDS region of the milk related gene was analyzed, and the correlation between the SNP loci and the daily milk yield was discovered in Yili. The active region of horse PRL and PRLR gene promoter was preliminarily explored, which provided a basis for the efficient breeding of Chinese milk horse and the preservation and utilization of germplasm resources.

【學(xué)位授予單位】：西北農(nóng)林科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S821

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