Screening of Interacting Proteins with Fungal Elicitor PevD1 by Yeast Two Hybrid System and High Expression of Recombinant in E. coli

Abstract

Abstract: PevD1, a fungal protein elicitor, was secreted from Verticillium dahliae and could induce hypersensitive responses（HR）and systemic acquired resistances（SAR）in tobacco plant. In this study, we screened its interaction partners using yeast two hybrid system with Arabidopsis thaliana cDNA library. Three potential interacting proteins were obtained. Homologous genes NR1, NR2 and NR4 in Nicotiana tobacum were cloned. Yeast two-hybrid binding assays confirmed NR2 protein strangely binding with PevD1. The entire coding region of NR2 gene was cloned into the bacterial expression vectors pMAL-C2X. After IPTG induction and SDS-PAGE analysis showed that the target protein was expressed at a high level in bacterial cells.

Key words: Fungal elicitor PevD1, Interacting protein, Yeast two hybrid system, Protein expression

Tang Xiaoli, Wu Wenxian, Han Lei, Yang Xiufen. Screening of Interacting Proteins with Fungal Elicitor PevD1 by Yeast Two Hybrid System and High Expression of Recombinant in E. coli[J]. Biotechnology Bulletin, 2014, 0(10): 113-118.

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