吴嘉, 杨红玉, 乔菊香, 等. 拟南芥AtIDD4与3×Flag融合蛋白表达植株的构建及其鉴定[J]. 西南林业大学学报（自然科学）, 2019, 39(4): 89–95 . DOI: 10.11929/j.swfu.201812024
引用本文: 吴嘉, 杨红玉, 乔菊香, 等. 拟南芥AtIDD4与3×Flag融合蛋白表达植株的构建及其鉴定[J]. 西南林业大学学报（自然科学）, 2019, 39(4): 89–95 . DOI: 10.11929/j.swfu.201812024
shu
Jia Wu, Hongyu Yang, Juxiang Qiao, Guobin Zhang, Juncheng Chen, Qiong Huang, Yunyue Wang. Construction and Identification of AtIDD4 and 3×Flag Fusion Protein Expressing in Arabidopsis thaliana[J]. Journal of Southwest Forestry University, 2019, 39(4): 89-95. DOI: 10.11929/j.swfu.201812024
Citation: Jia Wu, Hongyu Yang, Juxiang Qiao, Guobin Zhang, Juncheng Chen, Qiong Huang, Yunyue Wang. Construction and Identification of AtIDD4 and 3×Flag Fusion Protein Expressing in Arabidopsis thaliana[J]. Journal of Southwest Forestry University, 2019, 39(4): 89-95. DOI: 10.11929/j.swfu.201812024
shu

拟南芥AtIDD4与3×Flag融合蛋白表达植株的构建及其鉴定

Construction and Identification of AtIDD4 and 3×Flag Fusion Protein Expressing in Arabidopsis thaliana

    摘要
  • 摘要: 为了利用染色质免疫共沉淀技术研究转录因子AtIDD4蛋白与下游DNA的互作，通过构建35S-AtIDD4CDS-3×Flag农杆菌原核表达载体，利用农杆菌花浸法将35S-AtIDD4CDS-3×Flag结构转入野生型拟南芥中，实现ATIDD4和3×Flag标签的共表达和超表达。结果表明：通过PCR扩增、DNA测序、实时荧光定量RT-PCR等方法对转基因突变体进行鉴定，3×Flag标签与AtIDD4共表达，同时AtIDD4基因的表达量显著提高。

     

    Abstract: In order to study an interaction between transcription factor AtIDD4 and downstream DNA using chromatin immunoprecipitation (ChIP) technique, the prokaryotic expression vector of Agrobacterium tumefaciens 35S-AtIDD4CDS-3×Flag was constructed in this paper. And this structure was transferred into wild-type Arabidopsis thaliana to achieve co-expression and overexpression of the ATIDD4 and 3×Flag tag. The results showed that the expression of AtIDD4 and 3×Flag tag were examined by the methods of PCR amplification, DNA sequencing and quantitative RT-PCR, the AtIDD4 and 3×Flag were co-expressed. And the expression level of AtIDD4 was dramatic increased in transgenic plants.

     

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