脂多糖刺激对马氏珠母贝血细胞DNA甲基化修饰的影响  

Effect of Genomic DNA Methylation after Lipopolysaccharide Stimulation on the Homcytes from Pinctada fucata martensii

作　　者：焦钰[1,2] 梁金婵 谷泽丰 张鹏飞 李榕娇[1] 罗少杰 JIAO Yu;LIANG Jin-chan;GU Ze-feng;ZHANG Peng-fei;LI Rong-jiao;LUO Shao-jie(Fisheries College,Guangdong Ocean University,Zhanjiang 524088,China;Guangdong Technology Research Center for Pearl Aquaculture and Process,Zhanjiang 524025,China)

机构地区：[1]广东海洋大学水产学院,广东湛江524088 [2]广东省珍珠养殖与加工工程技术研究中心,广东湛江524025

出　　处：《广东海洋大学学报》2020年第3期7-13,共7页Journal of Guangdong Ocean University

基　　金：国家自然科学基金(31672626);大学生创新创业训练项目(CXXL2015041)。

摘　　要：【目的】研究马氏珠母贝(Pinctada fucata martensii)血细胞基因组DNA的甲基化修饰情况及其在脂多糖(LPS)刺激后的变化,为探究甲基化修饰在珍珠贝免疫防御中的作用提供理论基础。【方法】运用甲基化敏感扩增多态性法(MSAP)分析马氏珠母贝血细胞(正常养殖贝)以及注射LPS后(6、12、24 h)基因组CCGG位点甲基化的修饰水平变化,并对甲基化修饰位点进行回收测序。【结果】共获得1 102个清晰可辨的DNA条带,其中全甲基化位点条带215个,半甲基化位点条带127个,非甲基化位点条带760个。对照组血细胞基因组CCGG位点的甲基化修饰比例为23.76%,注射LPS后,甲基化比例为28.10%~48.25%,注射6 h时最低,12 h时最高;对特异性片段测序后,经Blast比对,得到5条存在甲基化修饰的基因序列,其中2条具有注释信息,分别是干扰素调控因子(Interferon regulatory factor)和磷脂酶BL2(Putative phospholipase B-like 2)。【结论】马氏珠母贝血细胞基因组CCGG位点的甲基化修饰参与调控LPS诱导的免疫反应,在免疫防御中发挥重要作用。【Objective】To analyze the DNA methylation level of the hemocytes from pearl oyster Pinctada fucata martensii after Lipopolysaccharide(LPS) stimulation, and explore the rules of DNA methylation in the immune defense in P. f. martensii. 【Methods】Methylation-sensitive amplification polymorphism(MSAP) technology was used to analyze the methylation levels of the hemocytes from P. f. martensii at 6 h, 12 h and 24 h after LPS injection. 【Results】A total of 1 102 clear and repetitive DNA bands, including 215 full-methylated sites, 127 hemi-methylated sites and 760 non-methylated sites were obtained. In the LPS group, the percentage of methylated sites was between 28.1% – 48.3%, and the lowest level was at 6 h, the highest level was at 12 h. After recovering and sequencing the specific bands, 5 DNA fragment were confirmed to have been methylated. Among them, two sequences can be annotated. The sequence were holologous to the interferon regulatory factor and a putative phospholipase B-like 2. 【Conclusion】 DNA methylation involved in the immune response stimulated by LPS, and it may play crucial roles in the immune defense of P. f. martensii.

关 键 词：马氏珠母贝 DNA甲基化 免疫防御 甲基化敏感扩增多态性法(MSAP) 

分 类 号：Q78[生物学—分子生物学] Q959.215.4