基于GWAS的6个免疫相关基因在大蒲莲猪和长白猪群体内的表达分析  

Expression Analysis of Six Immune-related Genes Selected from GWAS in Dapulian and Landrace Sus scrofa Population

作　　者：赵雪艳[1] 王彦平[1] 呼红梅[1] 王继英[1] 郭建凤[1] 

机构地区：[1]山东省农业科学院畜牧兽医研究所/山东省畜禽疫病防治与繁育重点实验室,济南250100

出　　处：《农业生物技术学报》2018年第2期263-271,共9页Journal of Agricultural Biotechnology

基　　金：山东省现代农业产业技术体系生猪创新团队建设项目(No.SDAIT-08-03);转基因生物新品种培育重大科技专项(No.2014ZX08009-045B);山东省农业科学院农业科技创新工程“畜禽种质资源保护与共享利用”项目(No.CXGC2016A04)

摘　　要：免疫相关基因的筛选与研究是猪(Sus scrofa)抗病育种的前提与基础。本研究基于课题组前期对仔猪的T淋巴细胞亚群指标进行的全基因组关联分析(genome-wide association study,GWAS),对5号染色体上一个1.05 Mb免疫调控区域内包含的6个免疫相关基因:白细胞分化抗原4(cluster of differentiation 4,CD4)、淋巴细胞活化基因-3(lymphocyte activation gene-3,LAG3)、白细胞分化抗原27(cluster of differentiation 27,CD27)、淋巴毒素β-受体(lymphotoxin beta receptor,LTBR)瘤坏死因子受体超家族成员1A(tumor necrosis factor receptor superfamily member 1a,TNFRSF1A)和白细胞分化抗原9(cluster of differentiation 9,CD9)在大蒲莲和长白仔猪群体外周血液中的表达量进行检测,并对各基因开展品种和母猪效应分析,为进一步筛选影响大蒲莲仔猪高抗病力的候选基因提供借鉴。采集大蒲莲猪(104头)和长白猪(171头)35日龄仔猪的外周血,提取总RNA,逆转录为cDNA后,通过qRT-PCR的方法进行上述免疫相关基因和内参基因β2-微球蛋白(beta-2-microglobulin,B_2M)的表达水平检测,在对各免疫基因表达量通过内参基因校正后,对矫正后的表达量ΔCt在大蒲莲猪和长白猪群体内的表达特征、品种和母猪效应进行分析。结果显示,大蒲莲仔猪CD4和CD27基因表达量极显著低于长白仔猪(P<0.01),且母猪效应差异也达到极显著水平(P<0.01);大蒲莲仔猪TNFRSF1A基因表达量显著低于长白仔猪(P<0.05);大蒲莲仔猪CD9基因表达量极显著高于长白仔猪(P<0.01)。本研究对基于GWAS的6个免疫相关基因在大蒲莲猪和长白猪群体内的表达开展分析,其中CD4、CD27、TNFRSF1A和CD9 4个基因在大蒲莲和长白仔猪群体间显著差异表达,这些基因可以考虑作为影响大蒲莲仔猪高抗病力的候选基因,为大蒲莲仔猪高抗病力性状的标记辅助选择研究提供基础。Screening immune-related genes is important for pig（Sus scrofa） disease resistant breeding. In ourprevious genome-wide association study（GWAS） of T lymphocyte subpopulation traits in piglets（Sus scrofa）,an immune regulation region of 1.05 Mb was identified, which contained 6 immune-related genes, that is,cluster of differentiation 4（CD4）, lymphocyte activation gene-3（LAG3）, cluster of differentiation（CD27）,lymphotoxin beta receptor（LTBR）, tumor necrosis factor receptor superfamily member 1 a（TNFRSF1 A） and cluster of differentiation（CD9）. In the present study, to unravel the 6 genes expression differences between breeds with different disease resistance, one typical Chinese indigenous breed（Dapulian） with higher disease resistance in North China and one commercial breed（Landrace） with lower disease resistance were used.Blood samples of clinically healthy Dapulian（n=104） and Landrace（n=171） piglets of 35 d old were collected, and their total RNA was extracted and reverse-transcribed into cDNA. Firstly, the mRNA expressions of these 6 genes and the reference gene beta-2-microglobulin（B2 M） were determined by quantitative real-time PCR（qRT-PCR）. Then, after normalization with reference gene B2 M, Six immunerelated genes expression characteristics and breed effects were performed analyses in the 2 breeds. The piglets used in this study were sampled from 13 Dapulian sows and 28 Landrace sows, finally, litter effect on these genes expression was also analyzed. The results of this study showed that the expressions of these 6 immunerelated genes in Dapulian and Landrace were lower than the reference gene（B2 M）, and individual difference in the population was small, with variation coefficients of 5.92~15.96 and 5.99~12.43 in Dapulian and Landrace respectively. Comparison between the 2 breeds, there were significant difference in the expression levels of CD4, CD27, TNFRSF1 A and CD9. Specifically, the expressions of CD4, CD27 and TNFRSF1 A in Dapulian piglets

关 键 词：大蒲莲猪 免疫相关基因 CD4 CD27 TNFRSF1A CD9 

分 类 号：S828.2[农业科学—畜牧学]