Construction and Identification of Bait Vectors with VP1 Gene of Encephalomyocarditis Virus in Yeast Two-hybrid System

doi:10.13560/j.cnki.biotech.bull.1985.2015.01.030

Abstract

Abstract: Bait vector pDHB1-VP1 was constructed for screening cellular proteins interacting with VP1 protein of encephalomyocarditis virus from yeast two-hybrid cDNA library of target cells in this study. VP1 gene was amplified and cloned into pMD18-T vector. After being verified by sequencing, it was directional cloning into bait vector pDHB1 of yeast two-hybrid system. Then the recombinant plasmid was identified by enzyme digestion and sequencing and transformed into yeast cells NMY51．The bait vectors’ expression and self-activation to reporter genes were tested．The results showed that the bait plasmid pDHB1-VP1 could express in yeast cells, and product size was 66 kD. It was specific binding with rabbit anti EMCV serum, which showed better immunogenicity. Bait plasmid pDHB1-VP1 was successfully constructed, could express in yeast cells and proved to be no self-activation to reporter genes．It could be used in the yeast two-hybrid system screening test．

Key words: encephalomyocarditis virus, vp1 protein, yeast two-hybrid, bait

Xie Jingying, Feng Ruofei, Xu Lei, Zhang Haixia, Li Xiangrong, Hou Lanxin, Ma Zhongren. Construction and Identification of Bait Vectors with VP1 Gene of Encephalomyocarditis Virus in Yeast Two-hybrid System[J]. Biotechnology Bulletin, 2015, 31(1): 198-202.

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