Yeast Plasmid Screen
Yeast Plasmid Screen - Related Videos
08:57
High-throughput Yeast Plasmid Overexpression Screen
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Cited by 12 •
2011
Here we describe a plasmid overexpression screen in Saccharomyces cerevisiae, using an arrayed plasmid library and a high-throughput yeast transformation protocol with a liquid handling robot.
11:59
Competitive Genomic Screens of Barcoded Yeast Libraries
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Cited by 44 •
2011
We have developed comprehensive, unbiased genome-wide screens to understand gene-drug and gene-environment interactions. Methods for screening these mutant collections are presented.
11:30
Unravelling the Function of a Bacterial Effector from a Non-cultivable Plant Pathogen Using a Yeast Two-hybrid Screen
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Cited by 8 •
2017
Bacterial effector proteins are important for establishing successful infections. This protocol describes the experimental identification of proteinaceous binding partners of a bacterial effector protein in its natural plant host. Identifying these effector interactions via yeast two-hybrid screens has become an important tool in unravelling molecular pathogenicity strategies.
08:30
Yeast Transformation and Cloning
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2023
S. cerevisiae are unicellular eukaryotes that are a commonly-used model organism in biological research. In the course of their work, yeast researchers rely upon the fundamental technique of transformation (the uptake of foreign DNA by the cell) to control gene expression, induce genetic deletions, express recombinant proteins, and label subcellular structures. This video provides an overview of how and why yeast transformation is carried out in the lab. The important features of yeast...
05:20
Yeast Two-Hybrid Assay to Determine Protein Self-Association in Yeast Cells
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2025
This video demonstrates the yeast two-hybrid assay to detect specific protein self-interactions. In this assay, the protein of interest is tagged to two domains of a transcription factor that encodes the β-galactosidase enzyme. When the two proteins interact, this causes transcription and translation to form the enzyme, which is then detected using a biochemical assay.
FAQs
Selection links cell growth to a plasmid-encoded marker. Under the chosen selective condition, yeast cells carrying the relevant plasmid can grow, while cells lacking that marker are not retained in the growing population. This enrichment step narrows the candidates before researchers examine colony appearance, reporter output, or plasmid content more closely.
A selectable marker primarily identifies cells that carry a plasmid under the applied growth condition. A reporter signal, colony phenotype, or other measurable trait provides additional evidence about the insert or biological activity. Using these outcomes separately or together helps distinguish simple plasmid presence from the function associated with its DNA content.
Molecular analysis examines the plasmid from a candidate colony to verify that the expected DNA insert is present. This adds information that growth or phenotype alone may not provide, because a colony can satisfy selection without confirming the intended plasmid structure or insert. The result strengthens interpretation of cloning and functional experiments.
The workflow begins with transformation, followed by growth under a condition that selects for a plasmid-encoded marker. Researchers then identify candidate colonies and evaluate them through colony phenotype, reporter signals, or molecular analysis. Comparing these observations with the desired insert or activity allows the screen to separate likely positives from other transformed cells.
In a plasmid library, many yeast cells may carry different plasmid contents. Selection first enriches cells whose plasmids provide the required marker, while colony-level or molecular readouts help identify candidates with a desired insert or biological activity. This combination makes the approach useful for examining numerous plasmid variants rather than checking only one cloning result.
The method supports cloning verification, gene-function assessment, functional genomics, and studies of protein interactions in living yeast cells. Its value comes from connecting plasmid content with growth, phenotype, reporter output, or another measurable result. In biology research, that connection helps relate a genetic construct to an observable cellular consequence.