大白猪肿瘤坏死因子α基因启动子区多态性及其对mRNA表达水平的影响  

Polymorphism of TNF-αGene Promoter and its Effect on mRNA Expression in Large White Pigs

作　　者：孙丽[1] 夏日炜 刘颖[1] 孙寿永[1] 包文斌[1] 吴圣龙[1] 

机构地区：[1]扬州大学动物科学与技术学院,江苏省动物遗传繁育与分子设计重点实验室,扬州225009

出　　处：《中国畜牧兽医》2015年第6期1518-1523,共6页China Animal Husbandry & Veterinary Medicine

基　　金：江苏省科技支撑计划(BE2013319、BE2014328);常州市科技支撑计划(CE2013011)

摘　　要：肿瘤坏死因子α(tumor necrosis factor,TNF-α)基因在机体固有免疫和炎症反应中发挥重要作用,本研究旨在分析大白猪TNF-α基因启动子区C-791T多态性及其与基因表达之间的关系,为分析TNF-α基因启动子区C-791T突变的遗传学效应提供科学依据。本试验利用PCR-SSCP方法对201头大白猪TNF-α基因启动子区C-791T位点多态性进行检测,同时利用Real-time PCR方法分析TNF-α基因C-791T位点不同基因型在大白猪11个组织中的mRNA差异表达情况。结果表明,大白猪TNF-α基因启动子区-791bp处存在碱基突变(C>T),并检测到3种基因型,分别为:CC、CT和TT。TNF-α基因在断奶仔猪脾脏、肺脏、胸腺和淋巴结等免疫器官中表达量均较高;TT和CT基因型个体在11个组织中TNF-α基因表达量普遍高于CC基因型个体,且在十二指肠及空肠组织中达到差异显著性水平(P<0.05)。因此,TNF-α基因启动子区C-791T突变对该基因的mRNA表达水平产生显著影响,有必要将TNF-α基因启动子区C-791T位点多态性作为提升机体肠道病原菌抗性的遗传标记进行深入研究。Tumor necrosis factor a （TNF-α） gene plays an important role in innate immunity and inflammatory responses,this study aimed at analyzing the relationship between the polymorphism of TNF-α gene promoter and gene expression in Large White pigs,and further providing a scien- tific basis for analyzing the genetic effect of TNF-α gene promoter C-791T mutation. This study detected the polymorphism at C-791T site of TNF-α gene promoter in 201 heads of Large White pigs by PCR-SSCP method. Meanwhile, the expression of TNF-α gene in 11 tissues was assayed by Real-time PCR and analyzed the differential expression among individuals with different geno- types at C-791T site. The results showed that there existed one mutation （C-791T） in TNF-α gene promoter and three genotypes were detected, which were CC, CT and TT. The mRNA expres- sions of TNF-α were higher in spleen, lung, thymus and lymph of weaned piglets. TNF-a gene expression levels of TT and CT genotypes individual were generally higher than CC genotype individual in 11 tis- sues, and the mRNA expression in TT and CT individuals were significant higher than that in CC individ- uals in the duodenum and jejunum tissue （P〈0.05）. Therefore, the C-791T mutation of TNF-α gene promoter indeed had a significant influence on the mRNA expression level of TNF-α gene, which indica-ted that it was necessary for us to carry out an in depth study of TNF-c~ gene promoter C-791T polymor- phism as genetic markers o{ enhancing the resistance to intestinal pathogenic bacteria.

关 键 词：猪 TNF-Α基因 多态性 差异表达 

分 类 号：S813.3[农业科学—畜牧学]