pGBKT7 DNA-BD Vector
pGBKT7 DNA-BD Vector
Brand: Takara Bio.
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pGBKT7 DNA-BD Vector
pGBKT7 is a yeast expression vector that is designed to express a fusion protein of the GAL4 DNA-binding domain (DNA-BD; amino acids 1–147) and a bait protein. The fusion protein is expressed at high levels from the medium ADH1 promoter. The fusion protein also contains a c-Myc epitope tag. To facilitate in vitro transcription/translation, pGBKT7 includes the T7 promoter between the GAL4 DNA-BD and the epitope tag.
Overview
- The most popular and widely cited yeast two-hybrid vectors
- pGBKT7 and pGADT7 are components of the Matchmaker Gold Yeast Two-Hybrid System
- pBridge expresses a third protein for modified two-hybrid studies
Applications
- Yeast two-hybrid screening
- Yeast three-hybrid screening
The yeast two-hybrid principle
The yeast two-hybrid principle. A bait protein interacts with the GAL4 recognition sequence (or promoter) upstream of a reporter gene. Transcription of the reporter is activated when a prey protein containing the GAL4 transcriptional activation domain interacts with the bait.
Matchmaker Gold reporter genes
Matchmaker Gold reporter genes. Yeast strain Y2HGold expresses 4 genes from 3 separate GAL4-responsive promoters in response to protein-protein interactions.
Tirode, F. et al. A conditionally expressed third partner stabilizes or prevents the formation of a transcriptional activator in a three-hybrid system. J. Biol. Chem. 272, 22995–22999 (1997).
Brachmann, R. and Boeke, J. Tag games in yeast: the two hybrid system and beyond. Curr. Opin. Biotechnol. 8, 561–568 (1997).
Osborne, M.A. et al. The yeast tribrid system—genetic detection of trans-phosphorylated ITAM-SH2-interactions. Biotechnology 13, 1474–1478 (1995).
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The yeast two-hybrid principle
The yeast two-hybrid principle. A bait protein interacts with the GAL4 recognition sequence (or promoter) upstream of a reporter gene. Transcription of the reporter is activated when a prey protein containing the GAL4 transcriptional activation domain interacts with the bait.
Matchmaker Gold reporter genes
Matchmaker Gold reporter genes. Yeast strain Y2HGold expresses 4 genes from 3 separate GAL4-responsive promoters in response to protein-protein interactions.
