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Yeast Signaling01:28

Yeast Signaling

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Yeasts are single-celled organisms, but unlike bacteria, they are eukaryotes (cells with a nucleus). Cell signaling in yeast is similar to signaling in other eukaryotic cells. A ligand, such as a protein or a small molecule released from a yeast cell, attaches to a receptor on the cell surface. The binding stimulates second-messenger kinases to activate or inactivate transcription factors that further regulate gene expression. Many of the yeast intracellular signaling cascades have similar...
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The mathematical expression known as the wave function, ψ, contains information about each orbital and the wavelike properties of electrons in an isolated atom. When atoms are bound together in a molecule, the wave functions combine to produce new mathematical descriptions that have different shapes. This process of combining the wave functions for atomic orbitals is called hybridization and is mathematically accomplished by the linear combination of atomic orbitals. The new orbitals that...
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Hybrid zones are narrow regions where two closely related species interact, mate, and produce hybrids. Relative to either parent species, hybrids may possess distinct phenotypic or genetic differences that impact their survival and reproductive success. The genetic variances introduced by hybridization influence species diversity and speciation processes within the hybrid zone.
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Related Experiment Video

Updated: Feb 8, 2026

Enhanced Yeast One-hybrid Screens To Identify Transcription Factor Binding To Human DNA Sequences
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Generating Yeast One-Hybrid DNA-Bait Strains.

John S Reece-Hoyes, Albertha J M Walhout

    Cold Spring Harbor Protocols
    |July 4, 2018
    PubMed
    Summary

    This protocol details generating DNA-bait strains for yeast one-hybrid (Y1H) screening. It outlines a three-step process using Gateway cloning for efficient DNA-bait integration into reporter vectors for genetic analysis.

    Area of Science:

    • Molecular Biology
    • Yeast Genetics
    • Biotechnology

    Background:

    • Yeast one-hybrid (Y1H) screening is a powerful method for identifying DNA-binding proteins and their interactions.
    • Efficient generation of DNA-bait strains is crucial for the success of Y1H screens.
    • Existing methods may lack versatility or efficiency for diverse DNA-bait constructs.

    Purpose of the Study:

    • To provide a detailed protocol for generating DNA-bait strains for Gateway-compatible Y1H screens.
    • To offer an alternative strategy for smaller DNA-baits using conventional ligation.
    • To enable the creation of versatile Entry clones for further applications.

    Main Methods:

    • Generating Entry clones with DNA-baits using Gateway cloning or primer annealing and ligation.

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  • Transferring DNA-baits into two Y1H reporter Destination vectors (pMW#2 [HIS3] and pMW#3 [LacZ]).
  • Integrating reporter constructs into the YM4271 yeast strain genome.

    • Main Results:

    • A comprehensive 3-step protocol for Y1H strain generation is presented.
    • The protocol accommodates both large (e.g., promoters) and small (≤100 bp) DNA-baits.
    • The generated Entry clones serve as versatile resources for various molecular biology applications.

    Conclusions:

    • This protocol facilitates the efficient and versatile generation of DNA-bait strains for Y1H screening.
    • The method supports the study of protein-DNA interactions and gene regulation.
    • The described approach enhances the utility of Y1H screening in genetic research.