文献类型：期刊文献

中文题名：AtMET1基因克隆及化学诱导表达分析

英文题名：At MET1 Gene Cloning and Chemical-inducible Expression Analysis

作者：梁立雄[1] 常英英[1] 高亚南[1] 王颜波[1] 张伟溪[1] 苏晓华[1] 张冰玉[1]

第一作者：梁立雄

机构：[1]林木遗传育种国家重点实验室,国家林业局林木培育重点实验室,中国林业科学研究院林业研究所,北京100091

年份：2016

卷号：29

期号：6

起止页码：946-950

中文期刊名：林业科学研究

外文期刊名：Forest Research

收录：CSTPCD;;Scopus;北大核心:【北大核心2014】；CSCD:【CSCD2015_2016】；

基金：国家高技术研究发展计划(863计划)课题(2013AA102703);林木遗传育种国家重点实验室基本科研业务费专项资金课题(TGB2013010)

语种：中文

中文关键词：pER8-MET1;17-β-雌二醇;化学诱导表达载体;瞬时表达

外文关键词：pER8-MET1; 17-β-estradiol; chemical-inducible expression vector; transient expression

分类号：S718.46

摘要：DNA甲基化是植物基因组中普遍存在的一种的重要表观遗传学机制,对植物生长发育及进化起着重要的调节作用[1-2]。植物体DNA甲基化的建立和维持受多个基因的协同调控,其中MET1（DN-MT1-like METHYLTRANSFERASE 1 gene）是在植物中最早分离出来的甲基化相关基因,编码DNA甲基化转移酶1（MET1,Methytransferase1）,该酶主要负责保持CG位点的甲基化,通过DNA甲基化作用影响植物的基因组结构、发育和进化。
[Objective]In order to study the function of Arabidopsis thaliana methyltransferase gene At MET1,17-β-estradiol inducible plant expression vector p ER8-MET1 was constructed,and its characters of inducible expression were detected. [Methods]The At MET1 gene from Arabidopsis thaliana was used as the target gene,and‘digestionligation＇method was applied for constructing the plant expression vector p ER8-MET1,which could be induced by17-β-estradiol. Then the vector was transferred to Agrobacterium tumefaciens LBA4404. The Agrobacterium strain containing p ER8-GFP vector was injected into the leaves of Nicotiana tabacum,then the leaves were induced by 17-β-estradiol in different levels of concentrations and time durations. QPCR was performed to detect GFP gene expression,through which the optimum concentration and time duration were screened. The Agrobacterium strain containing p ER8-MET1 vector was injected into the leaves of Nicotiana tabacum,then the leaves were induced by optimal concentration of 17-β-estradiol through different time durations. QPCR was performed to detect MET1 gene expression. [Results]The results of q PCR showed that 17-β-estradiol can effectively induce the expression of the target gene in p ER8-MET1 and the optimal concentration of 17-β-estradiol is 50 μmol · L- 1. The expression level of MET1 gene gradually increased with 17-β-estradiol treating time and reached the highest level at 12 h. [Conclusion]The p ER8-MET1 vector was successfully constructed. The construction of the plant expression vector p ER8-MET1 lays a good foundation for the mechanism study on the relationship between DNA methylation and phenotype variation.