禽呼肠孤病毒σA基因酵母双杂交诱饵载体的构建与鉴定

doi:10.16431/j.cnki.1671-7236.2017.03.032

Abstract

Abstract:

In order to screen the host protein that interacted with σA gene of avian reovirus, the bait vector pGBKT7-σA of avian reovirus σA gene in yeast two-hybrid system was constructed. Total RNA of avian reovirus S1133 standard strain was purified by using Trizol reagents. Avian reovirus σA gene was amplified by RT-PCR and was cloned into yeast two-hybrid bait vector pGBKT7. The recombinant bait vector was named as pGBKT7-σA. The recombinant bait vector was transformed into Saccharomyces cerevisiae Y2HGold. And then the recombinant bait vector of different concentrations was applied to the nutrient deficient medium. The results showed that the yeast two-hybrid bait vector had been constructed successfully, and the recombinant bait vector pGBKT7-σA on yeast cells had no toxicity and self-activation. This study laid a foundation for further screening host protein interacted with σA protein by yeast two-hybrid technique.

Key words: avian reovirus; σA gene; yeast two-hybrid system; bait vector

CLC Number:

S852.65

LU Heng, XIE Zhi-xun, XIE Li-ji, HUANG Li, HUANG Jiao-ling, WANG Sheng, ZHANG Yan-fang, ZENG Ting-ting, FAN Qing, LUO Si-si, XIE Zhi-qin, DENG Xian-wen. Construction and Identification of Bait Vector of Avian Reovirus σA Gene in Yeast Two-hybrid System[J]. , 2017, 44(3): 847-853.

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Construction and Identification of Bait Vector of Avian Reovirus σA Gene in Yeast Two-hybrid System

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