Yeast two-hybrid vectors

pGBKT7 is a yeast two-hybrid bait 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 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.

pGADT7 AD is a yeast two-hybrid expression vector that is designed to express a fusion protein between the GAL4 activation domain (AD; amino acids 768–881) and a protein of interest. The fusion protein is expressed at high levels from the full-length ADH1 promoter. The fusion protein also contains a hemagglutinin (HA) epitope tag. To facilitate in vitro transcription/translation, pGADT7 AD includes the T7 promoter between the GAL4 AD and the epitope tag.

pGBKT7 yeast two-hybrid bait vector

pGBKT7 is a yeast two-hybrid bait 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 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.

pGADT7 yeast two-hybrid prey vector

pGADT7 AD is a yeast two-hybrid expression vector that is designed to express a fusion protein between the GAL4 activation domain (AD; amino acids 768–881) and a protein of interest. The fusion protein is expressed at high levels from the full-length ADH1 promoter. The fusion protein also contains a hemagglutinin (HA) epitope tag. To facilitate in vitro transcription/translation, pGADT7 AD includes the T7 promoter between the GAL4 AD and the epitope tag.

pBridge three-hybrid bait vector

The pBridge Vector allows you to investigate tertiary protein complexes with any GAL4-based two-hybrid system. pBridge may be used instead of pGBKT7 in the Matchmaker Gold System. With its two distinct multiple cloning sites, pBridge allows expression of a DNA-BD fusion protein and another distinct protein that acts as the "bridge" between the bait and prey (Tirode et al 1997; Brachmann and Boeke 1997).

A three-hybrid system allows you to investigate more complex protein interactions than are detected by a standard two-hybrid analysis. The third protein in this system can participate in the interaction in several ways:

Acting as a bridge, by interacting with two proteins that do not directly interact with each other
Stabilizing a weak interaction between the two proteins
Modifying or inhibiting one or both of the proteins (Osbourne et al 1995)
Inhibiting the two-hybrid interaction

Expression of the additional protein is controlled by a conditional methionine promoter—it is expressed in the absence of methionine. This allows expression to be switched on or off by culturing in media with or without methionine. The effect of the third protein is indicated by expression of a reporter or nutritional marker. We offer yeast media lacking methionine (Cat. # 630430, 630429) that are ideal for use with this vector.

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Cat. #	Product	Size	Price
630404	pBridge Vector	20 ug	$453.00
This vector allows expression of two proteins: a DNA-binding domain fusion, and an additional protein under control of an inducible methionine promoter. In combination with an activation domain fusion and reporter strain from a GAL4-based two-hybrid system, one can establish a three hybrid system for the investigation of ternary protein complexes. Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents Components Image Data Back 1129.gif Back pBridge Three-Hybrid Vector map pBridge Three-Hybrid Vector map. A target protein is expressed as a fusion to the DNA-BD, while a second target protein is conditionally expressed from the P_met25 promoter. Interactions dependent on three separate proteins can be investigated when pBridge is combined with an AD fusion vector from a GAL4-based two-hybrid system.
630442	pGADT7 AD Vector	20 ug	$453.00
pGADT7 AD is a yeast expression vector that is designed to express a fusion protein between the GAL4 activation domain (AD; amino acids 768–881) and a protein of interest. The fusion protein is expressed at high levels from the full-length ADH1 promoter. The fusion protein also contains a hemagglutinin (HA) epitope tag. To facilitate in vitro transcription/translation, pGADT7 AD includes the T7 promoter between the GAL4 AD and the epitope tag. Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents Components Image Data Back 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. Back 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.
630443	pGBKT7 DNA-BD Vector	20 ug	$453.00
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. Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents Components Image Data Back 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. Back 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.

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

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Applications

Yeast two-hybrid screening
Yeast three-hybrid screening

References

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).

Additional product information

Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.

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