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  • ‹ Back to Matchmaker Gold yeast two-hybrid systems
  • Matchmaker Gold Yeast Two-Hybrid System
  • Make your own library for yeast two-hybrid screening
  • Mate and Plate yeast two-hybrid cDNA libraries
  • Aureobasidin A for improved selectable drug resistance in yeast
Learn more about our MatchMaker Gold Y2H systems. Matchmaker Gold yeast two-hybrid systems
Yeast two-hybrid products Yeast two-hybrid products
Yeast culture media including YPD, YPDA, minimal media, and amino acid dropout mixes Yeast media
Quick and easy yeast transformation kit Quick & easy yeast transformation
Home › Learning centers › Protein research › Matchmaker Gold yeast two-hybrid systems › Matchmaker Gold Yeast Two-Hybrid System

Protein research

  • Capturem technology
    • Capturem protocols
    • Capturem tech notes and applications
    • FAQs about Capturem technology
    • Capturem technology citations
    • Capturem posters
  • Antibody immunoprecipitation
    • IP and Co-IP of cardiac voltage-gated ion channel proteins
    • Tech note: thiophilic antibody purification resins
    • Tech note: IP and Co-IP
  • His-tag purification
    • Purification methods overview
    • TALON resin selection guide
    • Selection guide: His60 resin
    • xTractor Buffer is optimized for superior protein yield
    • Why tag a protein?
    • Tech note: cobalt resin
    • Simplified purification of active, secreted his-tagged proteins
    • Overview: His60
    • Tech note: Capturem technology
    • Tech note: Capturem large volume
    • Magnetic beads
    • FAQs: TALON
    • Protocols
      • Video: Capturem his maxiprep
      • Video: Capturem his miniprep
      • Visual protocol: Capturem his maxiprep
      • Visual protocol: Capturem his miniprep
      • Capturem nickel column reagent compatibility
      • TALON reagent compatibility
      • His60 reagent compatibility
      • TALON: Native vs denaturing purification
      • Protocol: denaturing purification with TALON resin, imidazole elution
      • Protocol: native purification with TALON resin, imidazole elution
      • Protocol: native purification with TALON resin, pH elution
  • Other tag purification
    • Streptavidin-based enrichment using Capturem technology
    • Selection guide: peptide tags
    • Myc-tagged protein purification overview
    • GST-tagged protein purification overview
  • Phosphoprotein and glycoprotein purification
    • Non-tagged protein purification overview
    • Phosphoprotein purification overview
  • Matchmaker Gold yeast two-hybrid systems
    • Matchmaker Gold Yeast Two-Hybrid System
    • Make your own library for yeast two-hybrid screening
    • Mate and Plate yeast two-hybrid cDNA libraries
    • Aureobasidin A for improved selectable drug resistance in yeast
  • Expression systems
    • Protein expression overview
    • Insect expression overview
    • Mammalian expression overview
    • pHEK293 Ultra expression overview
    • OKT3 expression in mammalian cells
    • Bacterial expression overview
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Learn more about our MatchMaker Gold Y2H systems. Matchmaker Gold yeast two-hybrid systems
Yeast two-hybrid products Yeast two-hybrid products
Yeast culture media including YPD, YPDA, minimal media, and amino acid dropout mixes Yeast media
Quick and easy yeast transformation kit Quick & easy yeast transformation
Tech Note

Matchmaker Gold Yeast Two-Hybrid System

  • Four reporters and three promoters lead to fewer false positives
  • Interacting fusion proteins produce resistance to Aureobasidin A—a very potent yeast antibiotic
  • Antibiotic, nutritional, and blue/white selection for simple yet stringent screens
Introduction Advantages Application data Conclusion References

Introduction  

Our Matchmaker systems are highly advanced tools for identifying and characterizing novel protein-protein interactions (PPIs). Our latest and most powerful incarnation, the Matchmaker Gold Yeast Two-Hybrid System, adds a sensitive Aureobasidin A (AbA; Takesako et al. 1991) antibiotic resistance marker to two nutritional reporters and blue/white color selection. This results in a four-reporter system with the easiest, most stringent yeast two-hybrid (Y2H) screening strategy available (Figure 1).

Aureobasidin A effectively kills yeast, but when the AUR1-C gene is turned on by a positive interaction between GAL4-hybrid proteins, the yeast become AbA-resistant. Secondary confirmation of positive clones employs four reporters regulated by three different GAL4-responsive promoters. This effectively eliminates false positives and leaves you with greater numbers of genuine positives. Quality screening results like these, as well as our simple Mate & Plate library screening protocol, save you time and make your search for PPIs faster, easier, and more fruitful.

Yeast two-hybrid system design

Figure 1. Yeast two-hybrid system design. Library-derived, transcription-activating prey fusion proteins that interact with the DNA-binding bait fusion protein activate the expression of reporter genes.

Y2H systems exploit the modular nature of eukaryotic transcription factors, which consist of a sequence-specific DNA-binding domain (DNA-BD) and an RNA Pol II-recruiting transcription activation domain (AD; Fields and Song 1989; Chien et al. 1991). In Matchmaker systems, a known protein of interest is fused to the DNA-BD of the yeast GAL4 transcription factor to create a “bait” protein. Interacting partner proteins, often derived from a library, are expressed as fusions to the AD of yeast GAL4, to create “prey” proteins (Figure 1). When pairs of interacting bait and prey fusion proteins are coexpressed in a yeast cell, GAL4 function is restored and the interacting fusion proteins are able to activate transcription of the reporter genes. In Matchmaker Gold, yeast clones that harbor interacting protein pairs can then be identified by the presence of the four reporters (Figure 2). In library screens, the plasmids containing the coding sequences for the library-derived prey proteins can be rescued from the surviving yeast clones and subjected to further analysis and sequencing.

Four reporters give Matchmaker Gold its high stringency

Figure 2. Four reporters give Matchmaker Gold its high stringency. Interacting bait and prey fusion proteins drive the expression of four different reporters from three different GAL4-responsive promoters (M1, G1, and G2), which are stably integrated in the genome of the reporter strain, Y2HGold. AbA resistance and the two auxotrophic reporters for histidine and adenine biosynthesis confer growth selection in the presence of AbA and on histidine- and adenine-deficient media, while the α-galactosidase reporter produces blue colonies in the presence of X-alpha-Gal.

Advantages  

Aureobasidin A selection eliminates background

Matchmaker Gold systems are unique because they use the AUR1-C gene as a novel reporter that confers resistance to AbA, a potent antibiotic toxic to S. cerevisiae. Resistance to this highly stable antifungal depsipeptide (see technical note) allows straightforward Y2H library screening to be achieved without the optimization required when using nutritional selection alone. HIS3-based selection can produce high numbers of background colonies in primary screens, whereas AbA very effectively kills yeast cells not expressing the AUR1-C reporter. As a result, even low-stringency primary screens using AbA are quite definitive and produce a high percentage of genuinely positive clones, without the interference of background colonies.

Four reporters identify genuine positives

The stringency of Matchmaker Gold lies in the use of four selective reporter genes: AUR1-C, HIS3, ADE2, and MEL1 (α-galactosidase), whose expression is driven by three different GAL4-binding promoters (Figure 2). All of the reporter genes are stably integrated into the genome of the Y2HGold reporter strain. This strategic combination of four reporters virtually eliminates false positives, especially those arising from spurious GAL4 promoter-binding prey proteins, which might bind one promoter sequence but not all three.

Superior to nutritional selection alone

The use of auxotrophic reporters alone for Y2H screening often requires optimization steps, especially in the case of HIS3-based selection, to ensure that the growth conditions are sufficiently selective. The leakiness of HIS3 selection necessitates the use of 3-AT (a competitive inhibitor of the His3 protein), which must be titrated and included in the growth medium to suppress the growth of background colonies. With AbA selection and the two nutritional markers of Matchmaker Gold, growth selection is sufficiently stringent so as to make the use of 3-AT unnecessary.

Mate & Plate libraries and screening

Another major advantage of Matchmaker Gold is that we’ve replaced cumbersome library-scale yeast transformation with an easy Mate & Plate strategy that consists of combining the two haploid yeast strains that independently express the bait and prey fusion proteins (Figure 3). Y2HGold is a MATα strain which harbors the four integrated reporter genes and is transformed with a pGBKT7 plasmid to express the bait protein. The library prey strain, Y187, is a MATα strain and the ideal mating partner for Y2HGold.

We offer a variety of pretransformed Mate and Plate libraries in Y187 (see technical note). Alternatively, users can either create and transform their own library using the Make Your Own “Mate & Plate” Library System (see technical note), or transform a plasmid expressing a user-selected prey fusion protein. Combining the Y2HGold and Y187 strains for mating is very easy and results in diploid yeast that coexpress the desired combinations of the bait and prey proteins.

The Mate & Plate protocol

Figure 3. The Mate & Plate protocol. To screen a Matchmaker Mate & Plate library, an aliquot of the library in the Y187 strain (MATα) is simply mixed with a bait-expressing culture of the Y2HGold strain (MATα). The mated strains are cultured overnight and plated on selective agar medium (e.g., –Leu/–Trp + AbA + X-α-Gal).

Application data  

High numbers of confirmed positive clones

Due to the strong selective power of AbA resistance, first-round, low stringency screening selects only for blue, AbA-resistant colonies. Two-thirds of these putative positive clones are later confirmed by high-stringency screening, which selects for all four reporters. As a result, we now recommend low-stringency screening to generate a large pool of colonies, followed by high-stringency verification. This leads to more genuine positives and fewer false positives (Figure 4).

 Secondary Matchmaker Gold screening confirms high numbers of positive clones

Figure 4. Secondary Matchmaker Gold screening confirms high numbers of positive clones. A Y2HGold bait containing the POU domain from the mouse Oct-4 transcription factor (BD-POUmOct4) was used to screen the Mate & Plate Universal Mouse (Normalized) Library for Oct-4-binding proteins. 32 colonies from a low-stringency primary screen (DDO + AbA, 60 ng/ml + X-α-Gal) were selected and replated/patched onto fresh low-stringency medium (Panel A) and onto high-stringency medium (QDO + AbA, 60 ng/ml + X-α-Gal) (Panel B) to confirm the expression of all four Matchmaker Gold reporters. Of the 32 originally selected colonies, 25 were confirmed positive for the 4 reporters. DDO = Double dropout medium: SD/–Leu/–Trp. QDO = Quadruple dropout medium: SD/–Ade/–His/–Leu/–Trp.

Mouse Oct-4-binding proteins identified in Matchmaker Gold library screening
ProteinFunction
E2I/Ubc9 Small ubiquitin-related modifier (SUMO) enzyme involved in sumoylation of Oct-4; regulates Oct-4 stability
PIAS1 An E3 ligase protein inhibitor of activated STAT1; a potent inhibitor of Oct-4-mediated transcriptional activation
PSMB5 Proteasome beta5 subunit; may mediate the interaction of Oct-4 with proteasomes, which regulate cellular processes through protein degradation.

Aureobasidin A resistance is highly selective

To demonstrate the highly selective properties of AbA resistance, AbA was titrated against three diploid Y2HGold clones that each expressed different bait/prey protein pairs (Figure 5). A negative control strain that coexpressed a noninteracting protein pair (BD-POUmOct4 bait and an unmodified GAL4 AD protein, AD-null), was completely unable to grow in the presence of AbA concentrations above 40 ng/ml. When the BD-POUmOct4 bait was paired with a putatively interacting prey protein (AD-E2I), colonies were able to grow in the presence of 70–100 ng/ml AbA. This result suggests that these proteins interact well enough to activate AUR1-C expression and allow colony growth on AbA medium. In a positive control, the strongly interacting protein pair of DNA-BD-p53 and AD-SV40 large T antigen enabled the vast majority of colonies to grow in the presence of 100 ng/ml AbA—the highest concentration tested, indicating a very high level of AUR1-C expression.

Titration of Aureobasidin A for three different Matchmaker protein pairs

Figure 5. Titration of Aureobasidin A for three different Matchmaker protein pairs. Y2HGold yeast clones coexpressing one of three bait/prey fusion protein pairs: a negative control (BD-POUmOct4 + AD-null); a pair of putative interactors (BD-POUmOct4 + AD-E21); or a positive control (BD-p53 + AD-T-Ag), were grown on DDO agar (SD/–Trp/–Leu) in the presence of increasing concentrations of AbA. The relative ability of each strain to grow in the presence of AbA reflects the strength of the interactions between the bait and prey proteins. Libraries are usually screened in the presence of AbA at 60 ng/ml, which demonstrates definitive selection for interacting hybrids. POUmOct4 = POU domain from the mouse transcription factor, Oct-4.

Conclusion  

Our highly stringent four-reporter Matchmaker Gold Yeast Two-Hybrid System and our wide selection of tissue-specific, normalized, and universal Mate & Plate libraries offer the most convenient and advanced Y2H screening tools available. You can even construct your own library with the Make Your Own “Mate & Plate” Library System. Library screens can now be accomplished in less time and with greater confidence than ever before.

References  

Chien, C. T., Bartel, P. L., Sternglanz, R. & Fields, S. The two-hybrid system: a method to identify and clone genes for proteins that interact with a protein of interest. Proc. Natl. Acad. Sci. U. S. A. 88, 9578–82 (1991).

Fields, S. & Song, O. A novel genetic system to detect protein-protein interactions. Nature 340, 245–6 (1989).

Takesako, K. et al. Aureobasidins, new antifungal antibiotics. Taxonomy, fermentation, isolation, and properties. J. Antibiot. (Tokyo). 44, 919–24 (1991).

Related Products

Cat. # Product Size License Quantity Details
630489 Matchmaker® Gold Yeast Two-Hybrid System Each USD $1059.00

Complete GAL4-based two-hybrid system to detect protein interactions. Yeast strain Y2HGold facilitates the elimination of many false positives that arise when screening an activation domain (AD) fusion library, because it contains four separate reporter genes that can be used to select for protein interactions. The system is also characterized by low background due to the Aureobasidin A resistance marker.

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 You May Also Like Image Data

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The yeast two-hybrid principle

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
Matchmaker Gold reporter genes. Yeast strain Y2HGold expresses 4 genes from 3 separate GAL4-responsive promoters in response to protein-protein interactions.

Back

Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency

Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency
Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency. Interacting bait and prey fusion proteins drive the expression of four different reporters from three different GAL4-responsive promoters (M1, G1, and G2), which are stably integrated in the genome of the reporter strain, Y2H Gold. Aureobasidin A (AbA) resistance and the two auxotrophic reporters for histidine and adenine biosynthesis confer growth selection in the presence of AbA and on histidine- and adenine-deficient media, while the a-galactosidase reporter produces blue colonies in the presence of X-alpha-Gal.

Back

630489: Matchmaker Gold Yeast Two-Hybrid System

630489: Matchmaker Gold Yeast Two-Hybrid System
630482 Mate & Plate™ Library - Universal Mouse (Normalized) 2 x 1 mL USD $665.00

This yeast two-hybrid library was constructed from mouse cDNA that had been previously normalized to preferentially remove abundant cDNAs derived from high-copy-number mRNAs. The normalization process incorporates a Duplex-Specific Nuclease (DSN) treatment and SMART technology, and increases the representation of low-copy-number transcripts in the library. This reduces the number of clones that must be screened to identify positive interactions, and facilitates the identification and characterization of novel protein-protein interactions.

A universal mouse cDNA library transformed into yeast strain Y187. The library can be readily mated to a MATa GAL4 reporter strain, such as AH109 or Y2HGold.

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.

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Use of pretransformed libraries simplifies screening

Use of pretransformed libraries simplifies screening
Use of pretransformed libraries simplifies screening.

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Reduction in abundance of highly expressed gene transcripts following cDNA normalization

Reduction in abundance of highly expressed gene transcripts following cDNA normalization
Reduction in abundance of highly expressed gene transcripts following cDNA normalization. cDNA normalization reduces the abundance of highly expressed gene transcripts. Data are shown for genes from mixed tissues, before and after normalization, which exhibit greater than 10,000 Mean Fluorescence Units (MFU), representing over 7,000 genes. Approximately 3,300 genes show a significant reduction in intensity, and thus abundance, following normalization. Due to the large volume of data obtained, the median MFU was plotted for groups of 100 genes before and after normalization.

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DSN-Normalization reduces the amount of highly abundant transcripts

DSN-Normalization reduces the amount of highly abundant transcripts
DSN-Normalization reduces the amount of highly abundant transcripts. Normalized (Lanes N) and non-normalized (Lanes C) Human Universal cDNA samples (PCR products) were electrophoresed on a 1.5% agarose gel and transferred to Hybond-N membrane. PCR-amplified probes of GAPDH and beta-actin were labeled with 32P-dATP and hybridized to the membrane.

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630482: Mate & Plate Library - Universal Mouse (Normalized)

630482: Mate & Plate Library - Universal Mouse (Normalized)

Required Products

Cat. # Product Size Price License Quantity Details
630489 Matchmaker® Gold Yeast Two-Hybrid System Each USD $1059.00

Complete GAL4-based two-hybrid system to detect protein interactions. Yeast strain Y2HGold facilitates the elimination of many false positives that arise when screening an activation domain (AD) fusion library, because it contains four separate reporter genes that can be used to select for protein interactions. The system is also characterized by low background due to the Aureobasidin A resistance marker.

Documents Components You May Also Like Image Data

Back

The yeast two-hybrid principle

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
Matchmaker Gold reporter genes. Yeast strain Y2HGold expresses 4 genes from 3 separate GAL4-responsive promoters in response to protein-protein interactions.

Back

Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency

Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency
Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency. Interacting bait and prey fusion proteins drive the expression of four different reporters from three different GAL4-responsive promoters (M1, G1, and G2), which are stably integrated in the genome of the reporter strain, Y2H Gold. Aureobasidin A (AbA) resistance and the two auxotrophic reporters for histidine and adenine biosynthesis confer growth selection in the presence of AbA and on histidine- and adenine-deficient media, while the a-galactosidase reporter produces blue colonies in the presence of X-alpha-Gal.

Back

630489: Matchmaker Gold Yeast Two-Hybrid System

630489: Matchmaker Gold Yeast Two-Hybrid System
630490 Make Your Own "Mate & Plate™" Library System 5 Rxns USD $1635.00

Mate & Plate Libraries are by far the easiest libraries to screen for protein-protein interactions using a GAL4 yeast two-hybrid system. Several Mate & Plate Libraries are available ready-made from Takara Bio. For libraries that are not available, this system provides the necessary components and a simple, highly efficient method to make your own Mate & Plate Library using SMART technology and the highly efficient homologous recombination machinery of Saccharomyces cerevisiae.

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

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Library generation using n vivo recombination in S. cerevisiae. Mate & Plate Libraries are created via recombination between your cDNA and the Matchmaker prey vector pGADT7-Rec.

 Library generation using n vivo recombination in S. cerevisiae. Mate & Plate Libraries are created via recombination between your cDNA and the Matchmaker prey vector pGADT7-Rec.
Library generation using in vivo recombination in S. cerevisiae. Mate & Plate Libraries are created via recombination between your cDNA and the Matchmaker prey vector pGADT7-Rec. The homologues are generated by SMART cDNA synthesis. Colonies are pooled, mixed, and aliquoted into multiple vials. Each single 1 ml vial can be used for a two-hybrid screen.

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Mate & Plate libraries display broad insert representation

Mate & Plate libraries display broad insert representation
Mate & Plate libraries display broad insert representation. A human bone marrow library was made using the Make Your Own “Mate & Plate” Library System. Inserts from 15 randomly picked colonies were analyzed by yeast colony PCR using the Advantage 2 Polymerase Mix (Cat. No. 639201), and the Matchmaker AD LD-Insert Screening Amplimer Set (Cat. No. 630433). As seen in Lanes 1–15, every colony contained an insert of a different size. Lane M: 1 kb DNA ladder molecular weight marker.

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SMART cDNA synthesis generates cDNA ends with homology to pGADT7-Rec

SMART cDNA synthesis generates cDNA ends with homology to pGADT7-Rec
SMART cDNA synthesis generates cDNA ends with homology to pGADT7-Rec.

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630490: Make Your Own "Mate & Plate" Library System

630490: Make Your Own "Mate & Plate" Library System

Required Products

Cat. # Product Size Price License Quantity Details
630489 Matchmaker® Gold Yeast Two-Hybrid System Each USD $1059.00

Complete GAL4-based two-hybrid system to detect protein interactions. Yeast strain Y2HGold facilitates the elimination of many false positives that arise when screening an activation domain (AD) fusion library, because it contains four separate reporter genes that can be used to select for protein interactions. The system is also characterized by low background due to the Aureobasidin A resistance marker.

Documents Components You May Also Like Image Data

Back

The yeast two-hybrid principle

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
Matchmaker Gold reporter genes. Yeast strain Y2HGold expresses 4 genes from 3 separate GAL4-responsive promoters in response to protein-protein interactions.

Back

Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency

Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency
Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency. Interacting bait and prey fusion proteins drive the expression of four different reporters from three different GAL4-responsive promoters (M1, G1, and G2), which are stably integrated in the genome of the reporter strain, Y2H Gold. Aureobasidin A (AbA) resistance and the two auxotrophic reporters for histidine and adenine biosynthesis confer growth selection in the presence of AbA and on histidine- and adenine-deficient media, while the a-galactosidase reporter produces blue colonies in the presence of X-alpha-Gal.

Back

630489: Matchmaker Gold Yeast Two-Hybrid System

630489: Matchmaker Gold Yeast Two-Hybrid System
630494 Yeast Media Set 2 Each USD $324.00

Complete media sets for two-hybrid screening. Highest performing yeast media-hybrid system.

Package Contents: 

  • 2 pouches YPDA Broth
  • 1 pouch YPDA with Agar
  • 1 pouch SD/-Leu Broth
  • 1 pouch SD/-Leu with Agar
  • 1 pouch SD/-Trp Broth
  • 1 pouch SD/-Trp with Agar
  • 10 pouches SD/-Leu/-Trp with Agar
  • 1 pouches SD/-Ade/-His/-Leu/-Trp with Agar
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.

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Yeast media pouches

Yeast media pouches
Yeast media pouches. Each ready-to-go pouch provides a precise amount of premixed media and supplements to make 0.5 L of broth or agar media. Yeast Media Sets contain a complete assortment of mixes for preparing eight specialized broth and agar media, and are designed to complement our advanced Matchmaker Gold Systems. The Yeast Media Set 2 Plus includes Aureobasidin A and X-alpha-Gal, as well as media.

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630494: Yeast Media Set 2

630494: Yeast Media Set 2
630495 Yeast Media Set 2 Plus Each USD $692.00

The Yeast Media Set 2 Plus contains the media and supplements needed for yeast two-hybrid library screening with the Matchmaker Gold Yeast Two-Hybrid System (Cat. No. 630489). For the ready-to-go, preformulated pouches, just add water and autoclave according to the instructions provided in the Yeast Media Protocol-at-a-Glance: no measuring, mixing, or pH adjustments are required. Directions for the use of Aureobasidin A and X-alpha-Gal are provided in the Matchmaker Gold Yeast Two-Hybrid System User Manual.

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
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
Matchmaker Gold reporter genes. Yeast strain Y2HGold expresses 4 genes from 3 separate GAL4-responsive promoters in response to protein-protein interactions.

Back

Yeast media pouches

Yeast media pouches
Yeast media pouches. Each ready-to-go pouch provides a precise amount of premixed media and supplements to make 0.5 L of broth or agar media. Yeast Media Sets contain a complete assortment of mixes for preparing eight specialized broth and agar media, and are designed to complement our advanced Matchmaker Gold Systems. The Yeast Media Set 2 Plus includes Aureobasidin A and X-alpha-Gal, as well as media.
630466 Aureobasidin A 1 mg USD $237.00

Aureobasidin A (AbA) is a cyclic depsipeptide antibiotic, isolated from the filamentous fungus Aureobasidium pullulans R106, which is toxic to yeast at low concentrations (0.1–0.5 μg/ml). Sensitive species include Saccharomyces cerevisiae, Schizosaccharomyces pombe, Candida glabrata, Aspergillus nidulans and A. niger. AbA inhibits a yeast enzyme, inositol phosphorylceramide (IPC) synthase, whichis expressed from the AUR1 gene. Expression of a mutant gene, AUR1-C, in transformed yeast confers resistance to the drug.

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

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Aureobasidin A (AbA) is a cyclic depsipeptide antibiotic that inhibits a yeast enzyme, inositol phosphorylceramide (IPC) synthase, expressed from the AUR1 gene

Aureobasidin A (AbA) is a cyclic depsipeptide antibiotic that inhibits a yeast enzyme, inositol phosphorylceramide (IPC) synthase, expressed from the AUR1 gene
Aureobasidin A (AbA) is a cyclic depsipeptide antibiotic that inhibits a yeast enzyme, inositol phosphorylceramide (IPC) synthase, expressed from the AUR1 gene. Expression of a mutant gene, AUR1-C, in transformed yeast confers resistance to the drug. It is this gene that is used in Matchmaker Gold systems as a reporter for protein interactions.

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630466: Aureobasidin A

630466: Aureobasidin A
630462 X-alpha-Gal 100 mg USD $192.00

X-α-Gal is a chromogenic substrate used to detect α-galactosidase activity. In yeast, α-galactosidase is a secreted reporter that is expressed from the MEL1 gene in response to GAL4 activation. α-galactosidase accumulates in the media and hydrolyses X-α-Gal causing yeast colonies to turn blue. X-α-Gal can be used with Matchmaker GAL4-based products to confirm protein interactions.

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

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X-Alpha-Gal detects secreted alpha-galactosidase activity following a GAL4-based two-hybrid interactions in Y2HGold yeast patches and colonies

X-Alpha-Gal detects secreted alpha-galactosidase activity following a GAL4-based two-hybrid interactions in Y2HGold yeast patches and colonies
X-Alpha-Gal detects secreted alpha-galactosidase activity following a GAL4-based two-hybrid interactions in Y2HGold yeast patches and colonies.

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Yeast colonies that express alpha-galactosidase in response to a positive two-hybrid interaction turn blue when grown on media containing X-alpha-Gal

Yeast colonies that express alpha-galactosidase in response to a positive two-hybrid interaction turn blue when grown on media containing X-alpha-Gal
Yeast colonies that express alpha-galactosidase in response to a positive two-hybrid interaction turn blue when grown on media containing X-alpha-Gal.
630463 X-alpha-Gal 250 mg USD $331.00

X-α-Gal is a chromogenic substrate used to detect α-galactosidase activity. In yeast, α-galactosidase is a secreted reporter that is expressed from the MEL1 gene in response to GAL4 activation. α-galactosidase accumulates in the media and hydrolyses X-α-Gal causing yeast colonies to turn blue. X-α-Gal can be used with Matchmaker GAL4-based products to confirm protein interactions.

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X-Alpha-Gal detects secreted alpha-galactosidase activity following a GAL4-based two-hybrid interactions in Y2HGold yeast patches and colonies

X-Alpha-Gal detects secreted alpha-galactosidase activity following a GAL4-based two-hybrid interactions in Y2HGold yeast patches and colonies
X-Alpha-Gal detects secreted alpha-galactosidase activity following a GAL4-based two-hybrid interactions in Y2HGold yeast patches and colonies.

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Yeast colonies that express alpha-galactosidase in response to a positive two-hybrid interaction turn blue when grown on media containing X-alpha-Gal

Yeast colonies that express alpha-galactosidase in response to a positive two-hybrid interaction turn blue when grown on media containing X-alpha-Gal
Yeast colonies that express alpha-galactosidase in response to a positive two-hybrid interaction turn blue when grown on media containing X-alpha-Gal.

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