为研究禽流感病毒感染对宿主全基因组甲基化的影响,本试验使用荧光甲基化敏感扩增片段多态性方法(fluorescent labeled methyiation sensitive amplified polymorphism,F-MSAP)检测了SPF鸡心脏、肝脏、脾脏、肺脏、肾脏、气管上段、气管下段、胸腺、法氏囊9个不同组织,和感染A/Goose/Guangdong/1/96,H5N1,A/Duek/Anhui/1/2006(安徽),H5N12株禽流感病毒SPF鸡心脏和肺脏2种组织全基因组甲基化在cCGG位点的cC碱基对甲基化状态,并分别对比了2组感染试验组与SPF对照组全基因组甲基化差异。结果显示,SPF对照组的甲基化程度,9种组织的总甲基化比率均在53%左右,其中气管上段最低(50.00%),肝脏最高(57.33%)。在心脏中,2个感染组样品内甲基化比率、外甲基化比率和总甲基化比率均高于SPF鸡对照组;而在肺脏中,广东株感染组各甲基化比率均低于对照组,安徽株感染组均高于对照组。本研究初步揭示了禽流感病毒感染过程中HSN1对宿主(鸡)不同组织全基因组甲基化水平的影响,为后续家禽抗病性状的分子机制研究提供了理论依据。
To reveal the effect of the infection of avian influenza virus on the genome-wide DNA methylation level of the host,the DNA methylation levels and patterns of CCGG sites in genomes was analyzed among nine different tissues of SPF chicken, and the heart and lung of SPF chicken infected by A/Goose/Guangdong/1/96, HSN1, A/Duck/Anhui/1/2006 and HSN1, respectively. The differences of DNA methylation level were compared between Guangdong group and SPF group,Anhui group and SPF group, respectively. The result indicates that the degrees of DNA methylation are all approximately 53.22%, which the lowest degree is 50.00% in upper part of trachea,and the highest degree is 52.33%. In heart,both of Guangdong group and Anhui group are higher than SPF group in terms of the level of DNA methylation;in lung,the level of DNA methy- lation of Anhui group is higher than the SPF group, but the level of Guangdong group is lower than SPF group. These results demonstrated that there was difference in the DNA methylation level between the chicken infected by AIV and SPF chicken,which suggested that the infection of avian influenza virus (HSN1) may have effect on the DNA methylation level of healthy hosts.