中国荷斯坦牛金黄色葡萄球菌诱导型乳房炎CDH13基因甲基化分析

doi:10.16431/j.cnki.1671-7236.2018.07.025

摘要/Abstract

摘要：

为探索钙黏蛋白13基因（cadherin 13，CDH13）甲基化在奶牛金黄色葡萄球菌乳房炎中的调控机制，本试验在已建立的金黄色葡萄球菌诱导型荷斯坦牛乳房炎模型的基础上利用重亚硫酸盐测序技术（bisulfite sequencing PCR，BSP）检测正常乳腺组织和乳房炎乳腺组织候选基因CDH13的甲基化程度，并分析其甲基化程度与基因表达水平的关系。结果显示，除金黄色葡萄球菌致病组（试验组）第12个CpG岛外，其余所有CpG岛均呈现不同程度的甲基化：-259处甲基化水平最高（50%~60%），-144、-135和-126处甲基化均较低，为5%左右。试验组和对照组总体甲基化率分别为10.13%±1.81%和14.43%±0.55%，不同位点和总体甲基化率无显著差异（P>0.05）。与正常乳腺组织相比，乳房炎感染组乳腺组织CDH13基因表达上调，CDH13基因-162和-93两个CpG岛甲基化水平与其基因表达呈显著负相关（P<0.05）。本试验结果表明，CDH13基因甲基化影响其基因表达，从而影响乳房炎的发生，为进一步探索其在金黄色葡萄球菌致病型乳房炎中的发病机制提供了参考依据。

关键词: CDH13基因; 乳房炎; 甲基化; 中国荷斯坦牛

Abstract:

In order to explore the regulatory mechanism of cadherin 13 (CDH13) gene methylation in mastitis caused by Staphylococcus aureus in dairy cows,the CDH13 gene methylation levels of healthy and mastitis infected mammary tissue were tested by bisulfite sequencing PCR (BSP),and the relationship between the level of CDH13 gene methylation and expression was analyzed.The results showed that,all of the CpG islands except the twelfth CpG of experimental group showed different levels of methylation.The methylation level was the highest in -259 (50% to 60%),and those at -144,-135 and -126 were low,which were all about 5%.The overall methylation level of experimen and control groups ware 10.13%±1.81% and 14.43%±0.55%,respectively,and there was no significant difference between different loci and overall methylation rate of this two groups (P>0.05).Compared with the healthy tissue,the expression of CDH13 gene in mammary tissue which were inflected by mastitis was up-regulation,the methylation level of -162 and -93 CpG islands were significantly negatively correlated with gene expression (P<0.05).The methylation of CDH13 gene affected its expression,thus affecting the occurrence of mastitis.These provided a reference for further exploring the pathogenesis of Staphylococcus aureus pathogenic mastitis.

Key words: CDH13 gene; mastitis; methylation; Chinese Holstein

中图分类号:

S858.23

王梦琦, 倪炜, 唐程, 郭佳禾, 张慧敏, 李明勋, 杨章平, 毛永江. 中国荷斯坦牛金黄色葡萄球菌诱导型乳房炎CDH13基因甲基化分析[J]. 《中国畜牧兽医》, 2018, 45(7): 1926-1932.

WANG Mengqi, NI Wei, TANG Cheng, GUO Jiahe, ZHANG Huimin, LI Mingxun, YANG Zhangping, MAO Yongjiang. Methylation Analysis of CDH13 Gene in Mastitis Induced by Staphylococcus aureus in Chinese Holstein[J]. , 2018, 45(7): 1926-1932.

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