仿刺参“化皮病”体壁组织DNA甲基化的MSAP分析

ANALYSIS OF DNA METHYLATION IN THE BODY WALL OF SEA CUCUMBER APOSTICHOPUS JAPONICUS WITH SKIN ULCERATION BY THE METHYLATION-SENSITIVE AMPLIFIED POLYMORPHISM(MSAP)

利用甲基化敏感扩增多态性(Methylation-sensitive amplified polymorphism,MSAP)技术分析了健康仿刺参(Apostichopus japonicus)体壁和"化皮病"仿刺参病变体壁、正常体壁DNA序列中CCGG位点的甲基化情况。结果显示,健康仿刺参体壁和"化皮病"仿刺参病变体壁、正常体壁总甲基化水平分别为(18.60±5.61)%、(26.70±6.82)%和(19.53±3.34)%,其中全甲基化水平分别为(13.97±4.86)%、(20.08±5.26)%和(15.42±2.61)%,半甲基化水平分别为(4.63±3.59)%、(6.62±3.80)%和(4.11±2.08)%。"化皮病"仿刺参病变体壁总甲基化水平和全甲基化水平显著高于健康仿刺参体壁和"化皮病"仿刺参正常体壁(P<0.05),健康仿刺参体壁与"化皮病"仿刺参正常体壁总甲基化水平和全甲基化水平差异不显著(P>0.05);三者的半甲基化水平无显著差异(P>0.05)。因此,推测仿刺参体壁"化皮"与DNA甲基化有关。

The technology of methylation-sensitive amplification polymorphism （MSAP） was used to investigate theDNA methylation patterns at CCGG sites in the body wall of the healthy and diseased Apostichopus japonicus. The results showed that the methylation levels of body wall of the healthy A. japonicus, ulcered and normal body wall of thediseased ones were （18.60±5.61）%, （26.70±6.82）% and （19.53±3.34）%, among which the full methylations were（13.97±4.86）%, （20.08±5.26）% and （15.42±2.61）%, and the hemi-methylations were （4.63±3.59）%, （6.62±3.80）% and（4.11±2.08）%, respectively. The total methylation and full methylation levels in ulcer body wall of diseased A. japonicus were obviously higher than those of normal body wall in both healthy and diseased A. japonicas that have no difference in the total methylation level and full methylation level in the body wall. There were no significant differences inthe hemimethylations among three groups. These results suggest that the ulceration of body wall may be associatedwith DNA methylation.