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Tech Note

Aureobasidin A: selectable drug resistance for yeast

  • Aureobasidin A resistance is a definitive selectable marker for yeast
  • Effectively reduces background in Matchmaker Gold yeast two-hybrid screens
  • Identifies genuine positives more easily, with fewer false positives

Introduction  

Aureobasidin A (AbA) is a potent and unique yeast antibiotic that kills S. cerevisiae at low concentrations (Takesako et. al. 1993). The drug is a cyclic multipeptide (Figure 1) that acts by inhibiting inositol phosphorylceramide synthase, an essential yeast enzyme. A mutant enzyme, encoded by the AUR1-C gene, confers resistance to AbA and can be used as a highly effective selectable marker that requires little to no optimization. Our optimized Matchmaker Gold Yeast Two-Hybrid System leverages this by including a reporter strain containing a stably-integrated AUR1-C gene, allowing for simple and rapid colony screening in yeast two-hybrid (Y2H) libraries using AbA selection.

Structure of Aureobasidin A

Figure 1. Structure of Aureobasidin A. Aureobasidin A (AbA; MW 1,100) is a cyclic depsipeptide antibiotic isolated from the fungus Aureobasidium pullulans R106. AbA inhibits the product of the yeast AUR1 gene (inositol phosphorylceramide synthase) and is toxic to S. cerevisiae at low concentrations (0.1 µg/ml). The gene product of a dominant mutant allele, AUR1-C, confers resistance to AbA and its expression can be used as a selectable marker.

Low background with AbA screening  

AbA selection virtually eliminates the high number of background colonies that often plague low-stringency primary screens that rely on nutritional markers alone (e.g., HIS3). Because AbA actually kills sensitive cells, rather than merely retarding their growth, AbA-based selection greatly favors the growth and identification of true positive clones. In general practice, a high percentage of clones that emerge from low-stringency primary screens using AbA selection are subsequently verified on high-stringency secondary screens that select for all four Matchmaker Gold reporters (AUR1-C, HIS3, ADE2, and MEL1).

AbA: ampicillin for yeast  

Many researchers have yearned for a yeast selection system similar to those used for E. coli or mammalian cells. In fact, AbA is used for yeast in a similar way as ampicillin and kanamycin are used for cloning in E. coli, or as G418 is used for selection of stably-transfected mammalian cell clones. AbA resistance is far easier to use in Y2H library screening than auxotrophic reporters, which often require additional optimization steps to achieve selective growth conditions.

If you wish to take your search for protein-protein interactions to the next level, look no further than AbA selection with the Matchmaker Gold Yeast Two-Hybrid System.

References  

Takesako, K. et al. Biological properties of aureobasidin A, a cyclic depsipeptide antifungal antibiotic. J. Antibiot. (Tokyo). 46, 1414–20 (1993).

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Cat. # Product Size Price License Quantity Details
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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.

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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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630490 Make Your Own "Mate & Plate™" Library System 5 Rxns $1413.00 *

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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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630489 Matchmaker® Gold Yeast Two-Hybrid System Each $914.00 *

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.

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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.

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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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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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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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630494 Yeast Media Set 2 Each $279.00 *

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
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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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630495 Yeast Media Set 2 Plus Each $585.00 *

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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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*You must be logged in to a Business Account in order to purchase these products online, since the purchase of these products may be restricted depending on your account type. Researchers at not-for-profit accounts receive a limited use license with their purchase of the product. Researchers at for-profit accounts must obtain a license prior to purchase. For details please contact licensing@takarabio.com.

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