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

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。

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 35 S-AtIDD4 CDS-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.