CAJ | 학술논문

目的：建立一种定量检测小鼠α-突触核蛋白基因（α-syn）内含子1甲基化水平的方法，分析不同脑区该区域甲基化水平的差异。方法使用MSPPrimer确定小鼠α-syn内含子1的CpG 岛区域。使用PyroMark assay design 2.0针对该CpG岛设计焦磷酸扩增及测序引物。采用焦磷酸测序法分析成年小鼠海马、皮质、纹状体及中脑四个脑区该区域甲基化水平。结果小鼠α-syn内含子1存在CpG岛。成功优化定量该CpG岛甲基化水平的焦磷酸测序方法。焦磷酸测序表明各脑区该区域甲基化均低于2％。结论建立了定量检测小鼠α-syn基因内含子1CpG岛甲基化水平的方法。该CpG岛甲基化对于成年小鼠各脑区中α-syn的表达不起主要调控作用。
목적：건립일충정량검측소서α-돌촉핵단백기인（α-syn）내함자1갑기화수평적방법，분석불동뇌구해구역갑기화수평적차이。방법사용MSPPrimer학정소서α-syn내함자1적CpG 도구역。사용PyroMark assay design 2.0침대해CpG도설계초린산확증급측서인물。채용초린산측서법분석성년소서해마、피질、문상체급중뇌사개뇌구해구역갑기화수평。결과소서α-syn내함자1존재CpG도。성공우화정량해CpG도갑기화수평적초린산측서방법。초린산측서표명각뇌구해구역갑기화균저우2％。결론건립료정량검측소서α-syn기인내함자1CpG도갑기화수평적방법。해CpG도갑기화대우성년소서각뇌구중α-syn적표체불기주요조공작용。
Objective To establish a quantitative method to examine the methylation level of CpG island located in the intron 1 of mouse α-synuclein, and analyze the difference of methylation in various brain regions .Methods CpG island in the intron 1 of mouse α-synuclein was identified using an online software MSPprimer .Pyrosequencing assay was designed using PyroMark assay design 2.0.DNA methylation was quantified in mouse hippocampus , cortex, striatum and midbrain using pyrosequencing assay .Results There was a CpG island in the intron 1 of mouse α-synuclein. Pyrosequencing assay measuring methylation level of this CpG island was optimized .It indicated that all of the brain regions were hypomethylated with a methylation rate of 2%or less.Conclusion DNA methylation does not play a major role in the transcription regulation of mouse α-synuclein in the brain .