番茄DNA甲基转移基因SlDRM2L的克隆及表达分析  

Cloning and expression analysis of tomato DNA methyltransferase SlDRM2L gene

作　　者：刘嘉荔 刘红达 汪宏涛 张玉 唐晓凤 LIU Jiali;LIU Hongda;WANG Hongtao;ZHANG Yu;TANG Xiaofeng(School of Food and Biological Engineering, Hefei University of Technology, Hefei 230601, China)

机构地区：[1]合肥工业大学食品与生物工程学院,安徽合肥230601

出　　处：《合肥工业大学学报（自然科学版）》2022年第3期406-410,共5页Journal of Hefei University of Technology：Natural Science

基　　金：国家自然科学基金资助项目(31671266)。

摘　　要：DNA甲基化主要是通过在甲基转移酶的催化下修饰基因组DNA来响应外界环境的胁迫,是表观遗传学的重要手段之一,参与植物中DNA甲基化起始的主要DNA甲基转移酶是DRM2。文章通过美国国家生物技术信息中心(National Center for Biotechnology Information,NCBI)数据库,在番茄中发现了一个与拟南芥胞嘧啶甲基转移酶DRM2同源性较高的序列,即SlDRM2L。通过GenBank(登录号NM-001246974)获得番茄DNA甲基转移酶SlDRM2L基因的cDNA全长序列。实时荧光定量聚合酶链式反应(polymerase chain reaction,PCR)结果表明SlDRM2L在所有组织中均有表达,叶和花的表达量最高。同时利用烟草瞬时表达系统将构建的绿色荧光瞬时表达载体35S::SlDRM2L-GFP用于亚细胞定位实验,通过激光共聚焦显微镜观察可知SlDRM2L蛋白定位于细胞核中;此外,在高温条件下(39℃,4 h),番茄SlDRM2L的基因转录水平明显提高,表明SlDRM2L介导的DNA甲基化可能受高温诱导。该文为发现SlDRM2L在植物生长发育中的作用提供了相关的科学依据,为深入研究SlDRM2L的功能奠定了理论基础。The DNA methylation catalyzed by DNA methyltransferase modifies genomic DNA to respond to stress of external environment,which is one of the important means of epigenetics.The main DNA methyltransferase involved in the initiation of DNA methylation in plants is DRM2.In this paper,through the National Center for Biotechnology Information(NCBI)database of USA,a sequence with high homology to Arabidopsis cytosine methyltransferase DRM2 was found,namely SlDRM2L.The full-length cDNA of tomato DNA methyltransferase SlDRM2L gene was obtained from GenBank(accession number NM-001246974).Quantitative real-time polymerase chain reaction(qRT-PCR)revealed that the expression of SlDRM2L was in all tissues of tomato,but higher expression in leaves and flowers.At the same time,the transient expression system of tobacco was used to construct the green fluorescent transient expression vector 35S::S1DRM2L-GFP for subcellular localization experiments.The observation through the laser confocal microscope revealed that SlDRM2L was expressed in the nucleus;In addition,SlDRM2L transcription level was significantly increased under high temperature(39℃,4 h),suggesting that SlDRM2L-mediated DNA methylation may be induced by high temperature.This paper provides relevant scientific basis for discovering the role of SlDRM2L in plant growth and development,and lays a theoretical foundation for in-depth study on the function of SlDRM2L.

关 键 词：番茄 SlDRM2L基因 高温诱导 亚细胞定位 实时荧光定量聚合酶链式反应(PCR) 

分 类 号：Q94-334[生物学—植物学] Q94-336