利用全基因组关联分析鉴定鸡肠炎沙门氏菌感染相关基因及其mRNA表达调控
发布时间：2018-11-22 18:54
【摘要】：肠炎沙门氏菌不仅是威胁家禽养殖业的一大危害性病原菌,而且是重要的食源性病原菌,容易造成食物中毒,引起人肠道疾病,危害着人类健康发展。畜禽产品、蛋类、肉制品是肠炎沙门氏菌重要的传染源,而且肠炎沙门氏菌有着广泛的传播途径,包括人类在内的广泛易感动物,是造成预防和控制肠炎沙门氏菌比较困难的重要原因。全基因组关联分析(GWAS)是在基因组范围内筛选影响遗传变异的高效能分析方法,具有统计效力高、精准度高等特点,并广泛应用于动物重要经济性状以及人类复杂疾病遗传学的研究。本研究通过肠炎沙门氏菌感染200只济宁百日鸡,采用case-cntrol实验设计,利用600K鸡高密度芯片对盲肠内沙门氏菌含量差异较大的济宁百日鸡进行全基因组关联分析(GWAS),在全基因组范围内筛选与肠炎沙门氏菌感染相关的基因或分子标记,通过实时定量PCR,进一步分析每个相关基因不同基因型的表达量与肠炎沙门氏菌感染的相关性,探讨肠炎沙门氏菌感染相关基因的表达调控。(1)通过GWAS分析,共检测到3个SNPs与肠炎沙门氏菌感染显著关联,分别位于3个不同的基因;其中rs313644723(C/T)位于1号染色上,基因Wnt7b(wingless-type MMTV integration site family)位于rs313644723(C/T)位点上游;rs80757564(G/T)位于5号染色体上,LRP5(low density lipoprotein receptor-related protein 5)基因第11段内含子上;rs313281555(A/G)位于9号染色体上LPP(LIM domain containing preferred translocation partner in lipoma)第7段内含子上。(2)差异SNPs位点不同基因型个体与盲肠沙门氏菌含菌量相关性分析结果:rs80757564(G/T)位点,GG基因型和TT、GT两基因型相比差异显著(P0.01),TT和GT两基因型之间没有显著差异;rs313281555(A/G)位点基因型之间,GG基因型和AA、AG两基因型之间有极显著差异,AA和AG两基因型之间没有显著差异;rs313644723(C/T)位点基因型之间,CC基因型和TT、CT基因型之间差异显著(P0.05),TT和CT基因型之间没有显著差异,且该位点与Tilquin等人研究白来航鸡感染沙门氏菌感染后泄殖腔内沙门氏菌含量相关的QTL(CLOAC-ST)相一致。(3)荧光定量PCR结果表明:在盲肠和脾脏组织中,对照组和实验组各候选基因不同基因型之间没有显著差异;而在不同基因型个体之间,在盲肠和脾脏组织中,LPP基因不同基因型之间没有显著差异;在脾脏组织中,基因Wnt7b,TT基因型显著高于CC基因型(P0.05);LRP5基因中,位点G15802980T的TT基因型和TG基因型的表达量极显著高于GG基因型(P0.01)。盲肠组织中Wnt7b各基因型之间具有显著差异(P0.05),而LRP5各基因型之间没有显著差异,但是两基因不同基因型之间具有相同的表达趋势。综上所述,通过GWAS筛选出的差异SNPs与肠炎沙门氏菌感染有重要作用,分别为:rs80757564(G/T);rs313281555(A/G)和rs313644723(C/T)。三个SNP所在基因LPP、Wnt7b与LRP5在鸡肠炎沙门氏菌感染中发挥表达调控作用
[Abstract]:Salmonella enteritidis is not only a harmful pathogen threatening poultry industry, but also an important foodborne pathogen, which can easily cause food poisoning, cause human intestinal diseases and endanger the healthy development of human beings. Livestock and poultry products, eggs and meat products are an important source of infection of Salmonella enteritis, and Salmonella enteritis has a wide range of transmission routes, including human beings, a wide range of susceptible animals, It is an important reason that it is difficult to prevent and control Salmonella enteritis. Genome-wide association analysis (GWAS) is a highly efficient method for screening genetic variation in the genome, which has the characteristics of high statistical efficacy and high accuracy. And widely used in animal important economic traits and human complex disease genetics research. In this study, 200 chickens infected with Salmonella enteritidis were infected with Jining 100 day chickens. The (GWAS), was analyzed by using case-cntrol design and 600K chicken high density microarray for the whole genome association analysis of Chicken with large difference in the content of Salmonella in the Cecum. The genes or molecular markers associated with Salmonella enteritis infection were screened in the whole genome, and the correlation between the expression of different genotypes of each related gene and the infection of Salmonella enteritidis was further analyzed by real-time quantitative PCR,. To investigate the expression and regulation of Salmonella enteritidis infection related genes. (1) by GWAS analysis, a total of three SNPs were found to be significantly associated with Salmonella enteritidis infection, respectively located in three different genes; Among them, rs313644723 (C / T) is located on 1 staining, Wnt7b (wingless-type MMTV integration site family) is upstream of rs313644723 (C / T), rs80757564 (G / T) is on chromosome 5, and LRP5 (low density lipoprotein receptor-related protein 5 is on intron 11. Rs313281555 (A / G) is located on the intron of segment 7 of LPP (LIM domain containing preferred translocation partner in lipoma) on chromosome 9. (2) the relationship between different genotypes of SNPs loci and the quantity of Salmonella cecum: rs80757564 (G / T) loci. There was no significant difference between GG genotype and TT,GT genotype (P0.01), TT and GT genotype). There were significant differences between GG genotype and AA,AG genotype among rs313281555 (A / G) genotypes, but there was no significant difference between AA and AG genotypes. There were significant differences between rs313644723 (C / T) locus genotypes, CC genotypes and TT,CT genotypes (P0.05) between), TT and CT genotypes. This locus is consistent with QTL (CLOAC-ST), which is related to the content of Salmonella in cloacal cavity after infection with Salmonella by Tilquin et al. (3) fluorescence quantitative PCR results showed that: in cecum and spleen tissues, There was no significant difference between different genotypes of candidate genes in control group and experimental group. However, there was no significant difference between different genotypes of LPP gene in the caecum and spleen, while in spleen, the genotype of Wnt7b,TT was significantly higher than that of CC (P0.05). In LRP5 gene, the expression of TT genotype and TG genotype at G15802980T was significantly higher than that of GG genotype (P0.01). There was significant difference among the genotypes of Wnt7b in cecum (P0.05), but there was no significant difference among the genotypes of LRP5, but there was the same trend of expression between different genotypes of the two genes. In conclusion, the difference between SNPs and Salmonella enteritidis infection was found by GWAS, which were rs80757564 (G / T), rs313281555 (A / G) and rs313644723 (C / T), respectively. Expression and Regulation of three SNP genes LPP,Wnt7b and LRP5 in Chicken Salmonella enteritidis infection
【学位授予单位】：山东农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S858.31
本文编号：2350241
[Abstract]:Salmonella enteritidis is not only a harmful pathogen threatening poultry industry, but also an important foodborne pathogen, which can easily cause food poisoning, cause human intestinal diseases and endanger the healthy development of human beings. Livestock and poultry products, eggs and meat products are an important source of infection of Salmonella enteritis, and Salmonella enteritis has a wide range of transmission routes, including human beings, a wide range of susceptible animals, It is an important reason that it is difficult to prevent and control Salmonella enteritis. Genome-wide association analysis (GWAS) is a highly efficient method for screening genetic variation in the genome, which has the characteristics of high statistical efficacy and high accuracy. And widely used in animal important economic traits and human complex disease genetics research. In this study, 200 chickens infected with Salmonella enteritidis were infected with Jining 100 day chickens. The (GWAS), was analyzed by using case-cntrol design and 600K chicken high density microarray for the whole genome association analysis of Chicken with large difference in the content of Salmonella in the Cecum. The genes or molecular markers associated with Salmonella enteritis infection were screened in the whole genome, and the correlation between the expression of different genotypes of each related gene and the infection of Salmonella enteritidis was further analyzed by real-time quantitative PCR,. To investigate the expression and regulation of Salmonella enteritidis infection related genes. (1) by GWAS analysis, a total of three SNPs were found to be significantly associated with Salmonella enteritidis infection, respectively located in three different genes; Among them, rs313644723 (C / T) is located on 1 staining, Wnt7b (wingless-type MMTV integration site family) is upstream of rs313644723 (C / T), rs80757564 (G / T) is on chromosome 5, and LRP5 (low density lipoprotein receptor-related protein 5 is on intron 11. Rs313281555 (A / G) is located on the intron of segment 7 of LPP (LIM domain containing preferred translocation partner in lipoma) on chromosome 9. (2) the relationship between different genotypes of SNPs loci and the quantity of Salmonella cecum: rs80757564 (G / T) loci. There was no significant difference between GG genotype and TT,GT genotype (P0.01), TT and GT genotype). There were significant differences between GG genotype and AA,AG genotype among rs313281555 (A / G) genotypes, but there was no significant difference between AA and AG genotypes. There were significant differences between rs313644723 (C / T) locus genotypes, CC genotypes and TT,CT genotypes (P0.05) between), TT and CT genotypes. This locus is consistent with QTL (CLOAC-ST), which is related to the content of Salmonella in cloacal cavity after infection with Salmonella by Tilquin et al. (3) fluorescence quantitative PCR results showed that: in cecum and spleen tissues, There was no significant difference between different genotypes of candidate genes in control group and experimental group. However, there was no significant difference between different genotypes of LPP gene in the caecum and spleen, while in spleen, the genotype of Wnt7b,TT was significantly higher than that of CC (P0.05). In LRP5 gene, the expression of TT genotype and TG genotype at G15802980T was significantly higher than that of GG genotype (P0.01). There was significant difference among the genotypes of Wnt7b in cecum (P0.05), but there was no significant difference among the genotypes of LRP5, but there was the same trend of expression between different genotypes of the two genes. In conclusion, the difference between SNPs and Salmonella enteritidis infection was found by GWAS, which were rs80757564 (G / T), rs313281555 (A / G) and rs313644723 (C / T), respectively. Expression and Regulation of three SNP genes LPP,Wnt7b and LRP5 in Chicken Salmonella enteritidis infection
【学位授予单位】：山东农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S858.31
【参考文献】
相关期刊论文 前9条
1 刘璐;李建超;刘冉冉;李庆贺;文杰;常国斌;郑麦青;陈国宏;赵桂苹;;不同品种鸡对肠炎沙门氏菌人工感染初期免疫应答反应的差异[J];畜牧兽医学报;2015年06期
2 陈峰;柏建岭;赵杨;荀鹏程;;全基因组关联研究中的统计分析方法[J];中华流行病学杂志;2011年04期
3 尹定子;宋海云;;Wnt信号通路:调控机理和生物学意义[J];中国细胞生物学学报;2011年02期
4 韩建文;张学军;;全基因组关联研究现状[J];遗传;2011年01期
5 严卫丽;;复杂疾病全基因组关联研究进展——遗传统计分析[J];遗传;2008年05期
6 魏笑笑;王宝维;荆丽珍;;家禽抗病育种研究进展[J];家禽科学;2006年07期
7 黄银花,李宁,孙汉,黄路生;家禽数量性状基因座定位的研究进展[J];遗传;2001年06期
8 苏玉虹,熊远著,邓昌彦;猪的肉质性状基因定位研究进展[J];遗传;2000年05期
9 鲁绍雄,吴常信;动物育种方法的回顾与展望[J];国外畜牧科技;2000年01期
相关硕士学位论文 前2条
1 李建超;鸡肠炎沙门氏菌感染早期免疫指标变化及全基因组甲基化研究[D];扬州大学;2014年
2 张磊;鸡部分免疫性状全基因组关联分析研究[D];中国农业科学院;2012年
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