Chapter title |
Protein–Protein Interactions: Yeast Two-Hybrid System
|
---|---|
Chapter number | 14 |
Book title |
Bacterial Protein Secretion Systems
|
Published in |
Methods in molecular biology, July 2017
|
DOI | 10.1007/978-1-4939-7033-9_14 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7031-5, 978-1-4939-7033-9
|
Authors |
Lin, Jer-Sheng, Lai, Erh-Min, Jer-Sheng Lin, Erh-Min Lai |
Editors |
Laure Journet, Eric Cascales |
Abstract |
The yeast two-hybrid system is a powerful and commonly used genetic tool to investigate interactions between artificial fusion proteins inside the nucleus of yeast. Here we describe how to use the Matchmaker GAL4-based yeast two-hybrid system to detect the interaction of the Agrobacterium type VI secretion system (T6SS) sheath components TssB and TssC41. The bait and prey gene are expressed as a fusion to the GAL4 DNA-binding domain (DNA-BD) and GAL4 activation domain (AD, prey/library fusion protein) respectively. When bait and prey fusion proteins interact in yeast nucleus, the DNA-BD and AD are brought into proximity, thereby activating the transcription of reporter genes. This technology can be widely used to identify interacting partners, confirm suspected interactions, and define interacting domains. |
Mendeley readers
Geographical breakdown
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Demographic breakdown
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Student > Bachelor | 12 | 16% |
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Researcher | 6 | 8% |
Professor | 3 | 4% |
Other | 3 | 4% |
Unknown | 25 | 34% |
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Linguistics | 1 | 1% |
Other | 4 | 5% |
Unknown | 26 | 36% |