Dong Wenhua, Yin Xuemei, Sun Li, Wang Jing, Sun Shouyong, Zhu Guoqiang, Wu Shenglong, Bao Wenbin
Key Laboratory for Animal Genetics, Breeding, Reproduction and Molecular Design of Jiangsu Province, College of Animal Science and Technology, Yangzhou University, Yangzhou, Jiangsu, PR China.
College of Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu, PR China.
Gene. 2015 Nov 15;573(1):70-4. doi: 10.1016/j.gene.2015.07.026. Epub 2015 Jul 11.
Diarrhea and edematous disease are two major causes of mortality in postweaning piglets. These conditions lead to huge economic losses in the swine industry. Escherichia coli F18 is the primary causative agent of these two diseases. Transported associated with antigen processing (TAP) plays an important role in the immune response and the TAP1 gene could be an effective anti-E. coli F18 molecular marker in pigs. The aim of this study was to determine the correlation between TAP1 gene promoter CpG island methylation status and mRNA expression in piglets. In this study, bisulfite sequencing PCR (BSP) was used to detect the methylation status of the TAP1 gene promoter CpG islands and fluorescence quantitative PCR was used to detect TAP1 expression in the jejunum of Sutai piglets from birth to weaning age. The fragment of the TAP1 gene promoter region under investigation has no mutation, has 13 putative transcription factor binding sites containing 19 CpG sites, and may be important for regulation of gene expression. With increasing age, the overall methylation levels decreased, while the TAP1 expression levels increased, indicating a negative correlation between TAP1 expression and promoter methylation levels. Variance analysis showed significant differences in the methylation status of CpG_4, CpG_13 and CpG_15 among the different age groups (P<0.05). Our data indicate that TAP1 expression is increased by demethylation of promoter CpG islands, with CpG_4, CpG_13 and CpG_15 implicated as the critical regulatory sites.
腹泻和水肿病是断奶仔猪死亡的两大主要原因。这些病症给养猪业造成了巨大的经济损失。大肠杆菌F18是这两种疾病的主要病原体。与抗原加工相关的转运体(TAP)在免疫反应中起重要作用,TAP1基因可能是猪抗大肠杆菌F18的有效分子标记。本研究的目的是确定仔猪TAP1基因启动子CpG岛甲基化状态与mRNA表达之间的相关性。在本研究中,采用亚硫酸氢盐测序PCR(BSP)检测TAP1基因启动子CpG岛的甲基化状态,并用荧光定量PCR检测苏太仔猪从出生到断奶期空肠中TAP1的表达。所研究的TAP1基因启动子区域片段无突变,有13个推定的转录因子结合位点,包含19个CpG位点,可能对基因表达调控很重要。随着年龄增长,总体甲基化水平下降,而TAP1表达水平上升,表明TAP1表达与启动子甲基化水平呈负相关。方差分析显示不同年龄组之间CpG_4、CpG_13和CpG_15的甲基化状态存在显著差异(P<0.05)。我们的数据表明,启动子CpG岛去甲基化可增加TAP1表达,其中CpG_4、CpG_13和CpG_15被认为是关键调控位点。