CAJ | 학술논문

甲基化敏感的扩增多态性分析方法（MSAP）是在扩增片段长度多态性技术基础上建立起来的主要用来检测样品基因组 DNA 甲基化情况的一种方法，特别适用于全基因组范围内检测 CCGG 位点的胞嘧啶甲基化情况，在高等动物和许多植物上已广泛应用，但尚未有在昆虫中应用该方法的报道。从 DNA 提取、酶切基因组DNA 用量、酶切和接头连接时间、电泳图谱显影方法等方面对 MSAP 方法进行了改进，并在水稻重要害虫白背飞虱上得到成功应用。这为将来在其它昆虫中使用该方法研究基因组 DNA 甲基化情况提供了一定借鉴和指导价值。且通过比较白背飞虱雌雄性别间和长短翅型间的 MSAP 图谱，发现雌雄性别间和长短翅型间的基因组 DNA甲基化式样都存在明显的差异，说明 DNA 甲基化可能参与了性别和翅型分化的调节。
갑기화민감적확증다태성분석방법（MSAP）시재확증편단장도다태성기술기출상건립기래적주요용래검측양품기인조 DNA 갑기화정황적일충방법，특별괄용우전기인조범위내검측 CCGG 위점적포밀정갑기화정황，재고등동물화허다식물상이엄범응용，단상미유재곤충중응용해방법적보도。종 DNA 제취、매절기인조DNA 용량、매절화접두련접시간、전영도보현영방법등방면대 MSAP 방법진행료개진，병재수도중요해충백배비슬상득도성공응용。저위장래재기타곤충중사용해방법연구기인조 DNA 갑기화정황제공료일정차감화지도개치。차통과비교백배비슬자웅성별간화장단시형간적 MSAP 도보，발현자웅성별간화장단시형간적기인조 DNA갑기화식양도존재명현적차이，설명 DNA 갑기화가능삼여료성별화시형분화적조절。
The methylation-sensitive amplification polymorphism (MSAP)technique is an amplified frag-ment length polymorphism (AFLP)based method for detection of DNA methylation.It represents a relia-ble technique for the detection of cytosine methylation alterations at the site of CCGG in a genome-wide perspective.MSAP has been extensively applied in plants and some animals,but no application in in-sects has been reported yet.The method was improved from DNA extraction,the amount of genomic DNA used for restriction enzyme digestion,time for enzyme digestions and ligation of adapters,and electropho-resis imaging method.And the method was applied successfully in Sogatella furcifera (Horvath),an im-portant rice pest in China.It will provide some references and guiding values for the study of genome DNA methylation in other insects by using this method in the future.By comparing MSAP maps between male and female or between long wing and short wing of S.furcifera,it was found that there were obvious differences in DNA methylation patterns.This indicates that DNA methylation may be involved in gender and wings type differentiation.