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Bulletin of Botanical Research ›› 2015, Vol. 35 ›› Issue (5): 741-745.doi: 10.7525/j.issn.1673-5102.2015.05.017

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Construction and Identification of Bait Vector Containing BBTV nsp Gene in Yeast Two-hybrid System

WANG Xiang;ZHANG Xiu-Chun;YU Nai-Tong;LIANG Jie;ZHOU Peng;LIU Zhi-Xin*   

  1. Key Laboratory of Biology and Genetic Resources of Tropical Crops,Haikou 571101
  • Online:2015-09-15 Published:2015-11-20
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Abstract: The nuclear shuttle protein(NSP) which encoded by banana bunchy top virus(BBTV) DNA6 plays an important role in infection, replication and transportation of virus. In order to research the interaction of BBTV DNA6 and host banana protein, the yeast two-hybrid system were used for the assay. pGBKT7-nsp genes was amplified using two pairs of specific primers, and their PCR products were mixed and amplified to obtain the 1/4 DNA products which contained EcoRⅠ and BamHⅠ enzyme site sequence. And then the DNA was cloned to pGBKT7 of yeast two-hybrid system. After confirmation with sequence analysis, the plasmid was transformed into the yeast cell Y2H, and its toxicity and transcriptional activation were tested by color assay. The pGBKT7-nsp had no selfactivation, and non-toxic to yeast cells, and it was consistent with the requirements of yeast two hybrid bait plasmids for the next step in the experiment.

Key words: banana bunchy top virus(BBTV), nsp genes, yeast two-hybrid, bait vector, self-activation

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