【摘要】：從江香豬是我國(guó)稀有的優(yōu)良地方微型豬種,但香豬的生長(zhǎng)慢的調(diào)控機(jī)制不清楚。本文利用Illumina Porcine SNP60(60K)豬全基因組SNP芯片對(duì)從江香豬生長(zhǎng)性狀關(guān)聯(lián)SNP位點(diǎn)進(jìn)行全基因組關(guān)聯(lián)分析(GWAS),并結(jié)合MSAP檢測(cè)從核基因整體水平分析從江香豬的遺傳多態(tài)性,得到以下結(jié)論:1.應(yīng)用甲基化敏感擴(kuò)增多態(tài)性(MSAP)技術(shù),通過(guò)篩選的10對(duì)引物對(duì)60頭從江香豬個(gè)體血液基因組和肝臟DNA胞嘧啶甲基化模式和水平進(jìn)行評(píng)估,克隆差異片段比對(duì)分析�？偣驳玫�5642條條帶,從江香豬DNA的總甲基化水平為34.17%,全甲基化水平為14.84%,半甲基化水平為19.33%。分離克隆得到3條高體重群特有和3條低體重群特有的甲基化條帶。其中A1-A5位于基因CLSTN3(Calsyntenin-3)、磷酸酰肌醇蛋白聚糖6(glypican-6-like)、泛素連接酶E3(HERC3)、蛋白酪氨酸磷酸酶受體G(Protein-tyrosine phosphatase gamma,PTPRG)內(nèi)含子上,A6位于豬的UDP2C1基因第一個(gè)外顯子前端(GeneID:LOC100515222,序列號(hào)NC_010450)。2.應(yīng)用Illumina Porcine SNP 60芯片對(duì)120份香豬和可樂(lè)豬基因組的SNP進(jìn)行檢測(cè),經(jīng)過(guò)嚴(yán)格的質(zhì)量控制,結(jié)合體長(zhǎng)性狀進(jìn)行GWAS分析,得到15個(gè)與體長(zhǎng)關(guān)聯(lián)的SNP,挑選其中3個(gè)SNP位點(diǎn)INRA0011611、MARC0076145和H3GA0051838進(jìn)行AS-PCR驗(yàn)證,結(jié)果顯示芯片檢測(cè)結(jié)果與AS-PCR檢測(cè)結(jié)果相符。3.對(duì)3個(gè)生長(zhǎng)相關(guān)SNP位點(diǎn)對(duì)12月齡香豬98份,18月齡香豬123份,24月齡香豬96份,大白豬48份,關(guān)嶺豬18份,榮昌豬48份、糯谷豬30份、黔北黑豬23份、江口蘿卜豬18份和宗地花豬28份樣本進(jìn)行AS-PCR檢測(cè)。結(jié)果顯示:位點(diǎn)1在各豬群體中基因型無(wú)偏好。大樣本關(guān)聯(lián)分析顯示該點(diǎn)與24月齡香豬頭長(zhǎng)呈弱的正相關(guān)(0.2870.200)。位點(diǎn)2位于基因ALDH5A1第7內(nèi)含子中,香豬群體中主要以G等位基因?yàn)閮?yōu)勢(shì)等位基因,同時(shí)關(guān)聯(lián)分析發(fā)現(xiàn)該點(diǎn)與18月齡香豬和24月齡香豬頭長(zhǎng)呈弱的負(fù)相關(guān)(-0.201-0.200,-0.310-0.200)。位點(diǎn)3位于基因TCEAL4上游,大樣本驗(yàn)證結(jié)果顯示在香豬群體中以GG基因型為主,G等位基因?yàn)閮?yōu)勢(shì)等位基因。這些SNP位點(diǎn)功能的進(jìn)一步研究和了解,對(duì)保護(hù)從江香豬種質(zhì)資源,提高香豬養(yǎng)殖業(yè)發(fā)展速度具有一定借鑒作用。
[Abstract]:Congjiang Xiang pig is a rare and fine local miniature pig breed in China, but the regulation mechanism of its slow growth is not clear. In this paper, Illumina Porcine SNP60 (60K) porcine whole genome SNP chip was used to analyze the whole genome association SNP locus of Congjiang Xiang pig, and the genetic polymorphism of Congjiang Xiang pig was analyzed from the whole nuclear gene level with MSAP detection. The conclusions are as follows: 1. The methylation-sensitive amplified polymorphic (MSAP) (MSAP) technique was used to evaluate the methylation patterns and levels of DNA cytosine methylation in the blood and liver of 60 Congjiang pigs by screening 10 pairs of primers. A total of 5642 bands were obtained. The total methylation level of Congangxiang pig DNA was 34.17, the total methylation level was 14.84, and the semi-methylation level was 19.33. Three methylation bands specific to high body weight group and three low body weight groups were isolated and cloned. A1-A5 is located in intron of gene CLSTN3 (Calsyntenin-3), phosphoinositide proteoglycan 6 (glypican-6-like), ubiquitin ligase E3 (HERC3) and protein tyrosine phosphatase receptor G (Protein-tyrosine phosphatase gamma,PTPRG). A6 is located at the front end of the first exon of UDP2C1 gene (GeneID:LOC100515222, sequence number NC_010450). The genomic SNP of 120 fragrant pigs and cola pigs were detected by Illumina Porcine SNP 60 microarray. After strict quality control, the combined length traits were analyzed by GWAS, and 15 SNP, associated with body length were obtained, among which 3 SNP loci INRA0011611, were selected. MARC0076145 and H3GA0051838 are verified by AS-PCR. The results show that the results of chip detection are consistent with those of AS-PCR. 3. 3. Among them, 98 were 12 months old, 123 were 18 months old, 96 were 24 months old, 48 were big white pigs, 18 were Guan Ling pigs, 48 were Rongchang pigs, 30 were glutinous pigs, 23 were northern Guizhou black pigs. AS-PCR was performed on 18 radish pigs in Jiangkou and 28 in Zongduhua pigs. The results showed that locus 1 had no preference for genotype in all pig populations. Large sample correlation analysis showed that there was a weak positive correlation (0.2870.200) between this point and the head length of 24 months old fragrant pig. Locus 2 was located in intron 7 of gene ALDH5A1, and G allele was the dominant allele in Xiang pig population, and the correlation analysis showed that there was a weak negative correlation between G allele and head length (-0.201-0.200) in 18 month old Xiang pig and 24 month old Xiang pig. -0.310-0.200) Locus 3 was located upstream of gene TCEAL4. The results of large sample validation showed that GG genotype was dominant in Xiang pig population, and G allele was the dominant allele. The further study and understanding of the functions of these SNP loci can be used for reference in protecting the germplasm resources of Congjiang Xiang pigs and improving the development speed of Xiang pig breeding.
【學(xué)位授予單位】：貴州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類(lèi)號(hào)】：S828

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