Construction and Self-activated Detection of the Baits of HC-Pro, P3N-PIPO, CP and VPg from Sugarcane streak mosaic virus for Yeast Two Hybrid System

doi:10.7668/hbnxb.2016.01.014

Abstract

Abstract: Sugarcane streak mosaic virus (SCSMV) is one of the main pathogens causing mosaic syndrome on sugarcane.Yeast two hybrid system (Y2HS) is the main technique to investigate the mechanism between pathogen with host plant.In order to elucidate the interaction between the SCSMV and sugarcane by Y2HS,the coding sequences of HC-Pro, P3N-PIPO, CP and VPg from SCSMV were cloned into the pGBKT7,respectively,to generate 4 bait vectors,pGBKT7-HC-Pro,pGBKT7-P3N-PIPO,pGBKT7-CP and pGBKT7-VPg.Then the 4 bait vectors were transformed into the yeast strain Y2HGold,respectively.All the Y2HGold strains containing the bait vectors grew well on the SD/-Trp media as the control.It suggested that the proteins encoded by the baits were not toxic to yeast.All these 4 transformed yeast strains developed white but not blue plaques on the SD/-Trp/X-α-gal media.And they could not survive on the SD/-Leu,SD/-Trp/-Ade,SD/-Trp/-His or SD/-Trp/X-α-gal/AbA media,which indicating the bait proteins could not activate the report genes,such as MEL1,ADE2,HIS3 or AUR1-C.These 4 bait vectors constructed in this study could be used to screen the host proteins interacting with them to investigate the molecular mechanism and pathogenesis of SCSMV infection on sugarcane.

Key words: Sugarcane streak mosaic virus, HC-Pro, P3N-PIPO, CP, VPg, Bait vector, Yeast two hybrid system

CLC Number:

ZHAI Yushan, PENG Lei, YANG Yongqing, DENG Yuqing, CHENG Guangyuan, ZHENG Yanru, XU Jingsheng. Construction and Self-activated Detection of the Baits of HC-Pro, P3N-PIPO, CP and VPg from Sugarcane streak mosaic virus for Yeast Two Hybrid System[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2016, 31(1): 83-89. doi: 10.7668/hbnxb.2016.01.014.

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