滇陆猪Nramp1基因多态性对其在组织中表达的影响  

Effects of Nramp1 gene polymorphism on its expression in the tissues of Dianlu pig

作　　者：张斌 韩敏 相德才 刘韶娜 沙茜 张桂生 赵智勇 赵彦光 ZHANG Bin;HAN Min;XIANG De-cai;LIU Shao-na;SHA Qian;ZHANG Gui-sheng;ZHAO Zhi-yong;ZHAO Yan-guang(Yunnan Animal Science and Veterinary Institute,Kunming 650224,China;Luliang County Dianlu Pig Research Institute,Qujing,Yunnan 655600,China)

机构地区：[1]云南省畜牧兽医科学院,昆明650224 [2]陆良县滇陆猪研究所,云南曲靖655600

出　　处：《南方农业学报》2020年第1期202-208,共7页Journal of Southern Agriculture

基　　金：云南省重点研发计划项目(2018BB003);云南省应用基础研究计划项目(2017FD055,2018FD001);云南省畜牧兽医科学院基础研究项目(2019RW012)~~

摘　　要：【目的】明确自然抵抗相关巨噬细胞蛋白1基因(Nramp1)多态性对其在滇陆猪群体各组织中表达的影响,从分子遗传学角度阐述滇陆猪不同基因型个体对疫病免疫的遗传差异,为其抗病育种选育提供参考依据。【方法】采用PCR对27头滇陆猪Nramp1基因进行扩增,以DNASTAR 5.0进行序列比对分析及多态性(SNP)位点检测;同时利用实时荧光定量PCR检测Nramp1基因在滇陆猪各组织中的相对表达量,并运用SPSS 19.0分析Nramp1基因多态性对其在滇陆猪群体各组织中表达的影响。【结果】在滇陆猪Nramp1基因第5外显子、第5内含子和第6外显子上分别发现SNP1(T→C)、SNP2(A→G)和SNP3(G→A)等3个SNPs位点,都只存在2种基因型,且SNP1位点和SNP3位点在试验猪群体中属于低度多态[多态信息含量(PIC)/杂合度(He)<0.25)],SNP2位点属于中度多态(0.25肝脏>脾脏>心脏;此外,Nramp1基因在不同基因型滇陆猪群体中表现出的组织差异表达也不同,尤其是Nramp1基因在SNP2位点纯合群体肝脏中的相对表达量显著低于在杂合群体肝脏中的相对表达量,说明Nramp1基因多态性变异对其在滇陆猪组织中的表达有显著影响。【结论】Nramp1基因SNP位点变异能影响Nramp1基因在滇陆猪各组织中的表达变化,从而影响机体的免疫应答反应,其中SNP2位点可作为滇陆猪抗病育种的分子标记。【Objective】To clarify the effect of polymorphism of natural resistance-related macrophage protein 1 gene(Nramp1)on its expression in tissues of Dianlu pig,aiming at elaborating the genetic differences of different genotypes of Dianlu pigs on disease immunity from the perspective of molecular genetics,and providing a reference for disease resistance breeding.【Method】The Nramp1 gene of 27 Dianlu pigs were amplified by PCR,and the sequence alignment analysis and the polymorphic sites(SNP)were detected with DNASTAR 5.0.Meanwhile,the real-time quantitative fluorescence PCR(qPCR)was used to detect the relative expression of the Nramp1 gene in tissues of Dianlu pigs,and the SPSS19.0 was used to analyze the effect of Nramp1 gene polymorphism on its expression in the tissues of Dianlu pig.【Result】Three SNPs of SNP1(T→C),SNP2(A→G)and SNP3(G→A)were found on the 5 thexon,5 thintron and 6 thexon of the Nramp1 gene in Dianlu pig,respectively.There were only two genotypes in each polymorphism.The SNP1 and SNP3 were at low polymorphism[polymorphism information content(PIC)/heterozygosity(He)<0.25]and SNP2 was at moderate polymorphism(0.25liver>spleen>heart.In addition,the tissue-differential expression of the Nramp1 gene in different genotype groups of Dianlu pig was also different,especially the Nramp1 gene expression in the liver of homozygous group at SNP2 was significantly lower than that of heterozytous group,indicating that the variation of the Nramp1 gene polymorphism site had significant effect on its expression in the tissues of Dianlu pig.【Conclusion】The variation of the Nramp1 gene SNP can influence Nramp1 gene expression change in the tissues of Dianlu pig,thereby affecting the immune response reaction of Dianlu pig body.SNP2 site can be used as a molecular marker for disease resistance breeding in Dianlu pig.

关 键 词：滇陆猪 自然抵抗相关巨噬细胞蛋白1基因(Nramp1) 多态性位点(SNPs) 基因型 组织差异表达 

分 类 号：S828.89[农业科学—畜牧学]