AtMET1基因克隆及化学诱导表达分析  

At MET1 Gene Cloning and Chemical-inducible Expression Analysis

作　　者：梁立雄 常英英[1] 高亚南[1] 王颜波 张伟溪[1] 苏晓华[1] 张冰玉[1] LIANG Li-xiong CHANG Ying-ying GAO Ya-nan WANG Yan-bo ZHANG Wei-xi SU Xiao-hua ZHANG Bing-yu(State Key Laboratory of Tree Genetics and Breeding, Key Laboratory of Tree Breeding and Cultivation of State Forestry Administration, Research Institute of Forestry Chinese Academy of Forestry, Beijing 100091, China)

机构地区：[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

出　　处：《林业科学研究》2016年第6期946-950,共5页Forest Research

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)

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.

关 键 词：pER8-MET1 17-Β-雌二醇 化学诱导表达载体 瞬时表达 

分 类 号：S718.46[农业科学—林学]