刺参(Apostichopus japonicus)不同组织基因组甲基化状态MSAP分析

利用MSAP技术分析成体刺参的呼吸树、肠、肌肉和体壁等组织的基因组DNA甲基化水平,获得了四个组织基因组甲基化率.甲基化水平由高到低依次是呼吸树、体壁、肌肉和肠,分别是35.77％、33.51％、32.72％和28.0％,其中全甲基化位点各占19.46％、18.39％、19.18％和15.97％.统计学分析结果显示肠组织的甲基化水平与其它组织的差异显著(P＜0.05),其余三个组织间差异不显著(P＞0.05),但呼吸树与肌肉的半甲基化水平差异显著(P＜0.05).由MSAP分析差异位点克隆得到四个甲基化特异性片段,经测序分析功能未知,推测为刺参基因足非编码区或新功能基因序列.

MSAP ANALYSIS OF GENOME DNA METHYLATION IN DIFFERENT TISSUES OF APOSTICHOPUS JAPONICUS

Genomes of respiratory tree, intestine, muscle, body wall of sea cucumber Apostichopus japonicus were compared for the methylation level adopting methylation-sensitive amplification polymorphism technique (MSAP). Results show that the methylation probability of the four tissues from high to low were respiratory tree > body wall > muscle > intestine, as 35.77%, 33.51%, 32.72% and 28.0% in sequence, among which the full methylation took up 19.46%, 18.39%, 19.18% and 15.97% orderly. The statistic results showed that the intestine had significant difference with the other three tissues (P<0.05); and the difference of methylation between muscle, respiratory tree and body wall was not obvious (P>0.05), while there was significant difference between muscle and respiration tree in hemi-methylation (P<0.05). Four specific fragments were got through analysis of difference site in MSAP strips, and the functions of those fragments were unknown after sequencing and blasting. It was supposed that those sequences were Non-coding region or new functional genes of A. japonicus.