AtMET1基因克隆及化学诱导表达分析

梁立雄; 常英英; 高亚南; 王颜波; 张伟溪; 苏晓华; 张冰玉

AtMET1基因克隆及化学诱导表达分析

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
基金项目:
国家高技术研究发展计划（863计划）课题（2013AA102703）和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）。
中图分类号: S718.46

AtMET1 Gene Cloning and Chemical-inducible Expression Analysis

1.
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
CLC number: S718.46

- pER8-MET1
- / 17-β-雌二醇
- / 化学诱导表达载体
- / 瞬时表达
Abstract: [Objective] In order to study the function of Arabidopsis thaliana methyltransferase gene AtMET1, 17-β-estradiol inducible plant expression vector pER8-MET1 was constructed, and its characters of inducible expression were detected. [Methods] The AtMET1 gene from Arabidopsis thaliana was used as the target gene, and ‘digestion-ligation’ method was applied for constructing the plant expression vector pER8-MET1, which could be induced by 17-β-estradiol. Then the vector was transferred to Agrobacterium tumefaciens LBA4404. The Agrobacterium strain containing pER8-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 pER8-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 qPCR showed that 17-β-estradiol can effectively induce the expression of the target gene in pER8-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 12h. [Conclusion] The pER8-MET1 vector was successfully constructed. The construction of the plant expression vector pER8-MET1 lays a good foundation for the mechanism study on the relationship between DNA methylation and phenotype variation.
- pER8-MET1
- / 17-β-estradiol
- / chemical-inducible expression vector
- / transient expression

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AtMET1基因克隆及化学诱导表达分析

AtMET1 Gene Cloning and Chemical-inducible Expression Analysis

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AtMET1 Gene Cloning and Chemical-inducible Expression Analysis

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AtMET1基因克隆及化学诱导表达分析

AtMET1 Gene Cloning and Chemical-inducible Expression Analysis

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AtMET1基因克隆及化学诱导表达分析

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AtMET1 Gene Cloning and Chemical-inducible Expression Analysis

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