A novel, easy and rapid method for constructing yeast two-hybrid vectors using In-Fusion technology
| Autor: | Xuan Zhang, Chengwei Li, Ju Zhang, Xu Kedong, Liu Xia, Li Xiaoli, Zhangkun Wei, Guangxuan Tan, Yi Zhang, Yulu Wang, Deshui Yu, Liao Libing, Ran Chen, Han Xiaomeng, Kun Liu |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Cloning 030102 biochemistry & molecular biology Computer science Two-hybrid screening Genetic Vectors Mutagenesis (molecular biology technique) Computational biology Polymerase Chain Reaction General Biochemistry Genetics and Molecular Biology Yeast Reverse Genetics 03 medical and health sciences Restriction enzyme 030104 developmental biology Plasmid Two-Hybrid System Techniques Yeasts Multiple cloning site Escherichia coli Mutagenesis Site-Directed Vector (molecular biology) Cloning Molecular Biotechnology Plasmids |
| Zdroj: | BioTechniques. 64(5) |
| ISSN: | 1940-9818 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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