小麦去甲基化酶基因DME-5B克隆与鉴定
Cloning and Identification of Wheat Demethylase Gene DME-5B
朱明一 1李一政 1穆红梅 1郭尚敬1
作者信息
摘要
小麦DME蛋白和水稻中ROS1是一同类DNA去甲基化酶,ROS1对水稻糊粉层数有限制作用,DME在小麦中是否有相似功能未见报道.本研究采用同源克隆技术,在小麦中克隆出TraesDME-5B基因,并对其编码蛋白的理化性质及结构特征进行生物学特性分析,为研究去甲基化酶DME对小麦种子发育中糊粉层的影响进行初步探索.结果显示,TraesDME-5B全长5 862bp,编码1 953个氨基酸,预测DME-5B蛋白为疏水蛋白,该蛋白的二级结构主要含有α-螺旋、无规卷曲等,其中无规则卷曲所占比例为60.73％,是DME-5B二级结构中最主要的构成元件,并且三级结构与二级结构预测结果高度一致.保守结构域分析显示,DME家族在禾本科植物进化过程中含有RRM_DME、ENDO3c superfamily和Perm-CXXC 3个保守结构域.结果表明,TraesDME-5B是小麦中一类DNA去甲基化酶,能够发挥DNA去甲基化作用.
Abstract
Wheat DME protein and rice ROS1 are a DNA demethylase of the same kind.ROS1 has a limiting ef-fect on the number of aleurone layers in rice.Whether DME has a similar function in wheat has not been reported.In this study,the gene TraesDME-5B was cloned from wheat by homologous cloning technique,and the physico-chemical properties and structural characteristics of the protein encoded by TraesDME-5B were analyzed in order to study the effect of demethylase DME on the aleurone layer during wheat seed development.The results showed that the total length of TraesDME-5B was 5 862 bp,encoding 1 953 amino acids,and DME-5B was predicted to be hydrophobic protein.The secondary structure of TraesDME-5B mainly consisted of α-helixes and random coils,among which the latter accounted for 60.73％,playing the role of the most important component in the secondary structure of DME-5B,meanwhile the prediction results of tertiary structure and secondary structure were highly consistent.According to the conserved domains analysis,the DME family contains three conserved domains namely RRM DME,Endo3C superfamily and Perm-CXXC during the evolution of Gramineae.The results showed that TraesDME-5B is a DNA demethylase in wheat,and functions in DNA demethylation.
关键词
DNA去甲基化/小麦/同源克隆/Traes/DME-5BKey words
DNA demethylation/Wheat/Homologous cloning/TraesDME-5B引用本文复制引用
基金项目
山东省农业科技园区产业提升工程项目(2019YQ035)
出版年
2024
NETL
NSTL
万方数据