BioTechniques
(May 2018)
A novel, easy and rapid method for constructing yeast two-hybrid vectors using In-Fusion technology
Deshui Yu,
Libing Liao,
Ju Zhang,
Yi Zhang,
Kedong Xu,
Kun Liu,
Xiaoli Li,
Guangxuan Tan,
Ran Chen,
Yulu Wang,
Xia Liu,
Xuan Zhang,
Xiaomeng Han,
Zhangkun Wei,
Chengwei Li
Affiliations
Deshui Yu
1Key Laboratory of Plant Genetics & Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, PR China
Libing Liao
2Henan Key Laboratory of Crop Molecular Breeding & Biorector, Zhoukou, 466001, PR China
Ju Zhang
1Key Laboratory of Plant Genetics & Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, PR China
Yi Zhang
1Key Laboratory of Plant Genetics & Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, PR China
Kedong Xu
1Key Laboratory of Plant Genetics & Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, PR China
Kun Liu
1Key Laboratory of Plant Genetics & Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, PR China
Xiaoli Li
1Key Laboratory of Plant Genetics & Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, PR China
Guangxuan Tan
1Key Laboratory of Plant Genetics & Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, PR China
Ran Chen
1Key Laboratory of Plant Genetics & Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, PR China
Yulu Wang
1Key Laboratory of Plant Genetics & Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, PR China
Xia Liu
1Key Laboratory of Plant Genetics & Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, PR China
Xuan Zhang
1Key Laboratory of Plant Genetics & Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, PR China
Xiaomeng Han
1Key Laboratory of Plant Genetics & Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, PR China
Zhangkun Wei
1Key Laboratory of Plant Genetics & Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, PR China
Chengwei Li
1Key Laboratory of Plant Genetics & Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, PR China
DOI
https://doi.org/10.2144/btn-2018-0007
Journal volume & issue
Vol. 64,
no. 5
Abstract
Read online
Yeast two-hybrid systems are powerful tools for analyzing interactions between proteins. Vector construction is an essential step in yeast two-hybrid experiments, which require bait and prey plasmids. In this study, we modified the multiple cloning site sequence of the yeast plasmid pGADT7 by site-directed mutagenesis PCR to generate the pGADT7-In vector, which resulted in an easy and rapid method for constructing yeast two-hybrid vectors using the In-Fusion cloning technique. This method has three key advantages: only one pair of primers and one round of PCR are needed to generate bait and prey plasmids for each gene, it is restriction endonuclease- and ligase-independent, and it is fast and easily performed.
Keywords
WeChat QR code
Close
