Construction of a western flower thrip yeast two-hybrid library and TSWV membrane protein bait vectors
Author of the article:WAN Yan-Ran1** ZHENG Xiao-Bin2 YUAN Jiang-Jiang2 ZHANG You-Jun2 WU Qing-Jun2***
Author's Workplace：1. College of Plant Protection, Agricultural University of Hebei, Baoding 071000, China; 2. Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, China
Key Words： Frankliniella occidentalis; tomato spotted wilt virus; yeast two hybrid library; bait vector
Abstract：
[Objectives] To screen proteins involved
in the interaction between western flower thrips and the Tomato spotted wilt virus (TSWV). [Methods] A western flower thrip cDNA library and a
Y187 yeast library were constructed using the Make Your Own “Mate & Plate™”
Library System, and TSWV GN and GC bait vectors were constructed using the
Matchmaker Gold Yeast Two-Hybrid System. [Results] The capacity of the western flower thrip cDNA
library was 5.6×106 cfu，the average library recombination
rate was about 100%, and the average amplification sizes of insert fragments in
the cDNA library were above 1 kb. The transformation ratio and titer of the
Y187 yeast library were 5×106 cfu/μg and 2.97×108, respectively. The
constructed pGNKT7-GN and pGBKT7-GC bait vectors were well expressed in yeast
and had no self-activation and toxicity. [Conclusion] A western flower thrip yeast library and TSWV
membrane protein bait vectors were successfully constructed and can be used for
further hybrid library screening.