摘要
本研究分析奶牛健康乳腺组织和金黄色葡萄球菌感染的乳房炎乳腺组织中胶原蛋白Ⅰ型α2链基因(COL1A2)启动子区CpG岛甲基化与COL1A2基因表达的调控关系,为奶牛乳房炎的抗性育种及预防提供理论依据。利用生物信息学,分析COL1A2基因启动子区CpG岛分布及其转录因子结合位点;运用亚硫酸氢盐测序PCR(BSP)和RT-PCR法,分析健康乳腺组织和患乳房炎乳腺组织中,COL1A2基因启动子区的CpG岛甲基化程度与COL1A2基因表达及乳房炎的相关性。结果表明健康乳腺组织和患乳房炎乳腺组织中CpG岛甲基化差异不显著(P>0.05),均呈低甲基化状态(<50%),但位于转录因子SP1结合区域内的第4和第5 CpG位点,健康组甲基化程度(30%和60%)显著高于患乳房炎组(0和10%);健康组基因表达水平显著低于患乳房炎组(P<0.05)。说明,COL1A2基因在不同乳腺组织中的差异表达可能与其启动子区CpG岛转录因子SP1结合区域内的第4和第5CpG位点甲基化程度差异相关。
This study was to investigate the methylation pattern of COL1A2 gene promoter and its effect on gene ex-pression between the healthy cow’ s mammary tissues （ n=3） and mastitic cow’s tissues （n=3） to provide clues for disease-resistant breeding and prevention of mastitis in dairy cows. The CpG islands and its transcription factors were analyzed by bioinformatics. The degree of methylation of a CpG island in COL1A2 gene core promoter region and the expression of COL1A2 mRNA were detected by the bi-sulfite sequencing PCR and RT-PCR. No obvious differ-ence （P〉0. 05） was detected in the degree of methylation of the CpG island in healthy cows and mastitic cows which＆amp;nbsp;both showed hypomethylation （〈50% of CpG sites on a given methylated strand）. The methylations at the 4th and 5th CpG sites within a SP1 binding site were higher in healthy mammary tissues（30% and 60%） than those in mastitic tissues（0 and 10%）. The expression of COL1A2 mRNA in healthy group was lower than that in mastitic group （P〈0. 05）. The dif-ferent degrees of methylation at the 4th and 5th CpG sites may contribute to the differential expression of the COL1A2 gene in mastitic and healthy mammary.
出处
《江苏农业学报》
CSCD
北大核心
2014年第3期574-580,共7页
Jiangsu Journal of Agricultural Sciences
基金
国家自然基金项目(31371255、31271328、31000543)
“十二五”国家科技支撑项目(2011BAD19B02-03)
“十二五”农业部转基因生物新品种培育重大专项子课题(2013ZX08007-001)
