Improve Research Reproducibility A Bio-protocol resource
Concise Method
tooltip

Y2H assay

CS Chen Sun
GY Gai-fang Yao
LL Li-xia Li
TL Ting-ting Li
YZ Yu-qi Zhao
KH Kang-di Hu
CZ Conghe Zhang
HZ Hua Zhang
request Request a Protocol
ask Ask a question
Favorite

Full-length WRKY71 sequences were cloned into pGBKT7 containing Nde I and Pst I for the transactivation activity assay. Full-length BRG3 was cloned into pGADT7 containing EcoR I and Xho I and co-transformed in pairs into yeast strain AH109 cells using the LiAC-PEG method. The primers used are presented in Supplemental Table S5. Selection of co-transformed colonies was performed on SD(-Leu/-Trp) growth medium. Protein interactions were examined via growth on SD(-Leu/-Trp/-His/-Ade) screening medium. Co-transformations of pGADT7-T and pGBKT7-53/pGBKT7-Lam were used as the positive and negative control, respectively (Li et al. 2020a, 2020b).

Copyright and License information:  ©2023-02-03 American Society of Plant Biologists 2023. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com
This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/pages/standard-publication-reuse-rights)

Do you have any questions about this protocol?

Post your question to gather feedback from the community. We will also invite the authors of this article to respond.

0/150

tip Tips for asking effective questions

+ Description

Write a detailed description. Include all information that will help others answer your question including experimental processes, conditions, and relevant images.

post Post a Question
0 Q&A