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Yeast One-Hybrid (Y1H) Assay

XW Xuxu Wang
XC Xiude Chen
QW Qingjie Wang
MC Min Chen
XL Xiao Liu
DG Dongsheng Gao
DL Dongmei Li
LL Ling Li
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To conduct the Y1H assay, we first cloned the open reading frames of MdBZR1 and MdBZR1-2like into the pGADT7 vector. The promoters of MdGA20ox2 and MdGA3ox1 were cloned into the pHIS2 vector with their native promoters. We co-transformed four pairs of constructs, namely pGADT7-MdBZR1/pHIS-MdGA20ox2, pGADT7-MdBZR1/pHIS-MdGA3ox1, pGADT7-MdBZR1-2like/pHIS-MdGA20ox2, and pGADT7-MdBZR1-2like/pHIS-MdGA3ox1, into yeast strain Y187 according to the manufacturer’s instructions (Clontech, Beijing, China). The transformed yeast strains were grown on SD/-Leu/-Trp and SD/-Leu/-Trp/-His/260mM 3-amino-1,2,4-triazole (3-AT) plates.

Copyright and License information:  ©2019 Wang, Chen, Wang, Chen, Liu, Gao, Li and Li
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