Yeast Electroporation
Yeast Electroporation - Related Videos
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...
12:19
Bacterial Transformation: Electroporation
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2023
The term “transformation” refers cellular ingestion of foreign DNA. In nature, transformation can occur in certain types of bacteria. In molecular biology, however, transformation is artificially induced through the creation of pores in the bacterial cell walls. Bacterial cells that are able to take up DNA from the environment are called competent cells. Electrocompetent cells can be produced in the laboratory and transformation of these cells can be achieve via the application of an...
08:30
Yeast Maintenance
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2023
Research performed in the yeast Saccharomyces cerevisiae has significantly improved our understanding of important cellular phenomona such as regulation of the cell cycle, aging, and cell death. The many benefits of working with S. cerevisiae include the facts that they are inexpensive to grow in the lab and that many ready-to-use strains are now commercially available. Nevertheless, proper maintenance of this organism is critical for successful experiments. This video will provide an overview...
07:48
Yeast Reproduction
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2023
Saccharomyces cerevisiae is a species of yeast that is an extremely valuable model organism. Importantly, S. cerevisiae is a unicellular eukaryote that undergoes many of the same biological processes as humans. This video provides an introduction to the yeast cell cycle, and explains how S. cerevisiae reproduces both asexually and sexually Yeast reproduce asexually through a process known as budding. In contrast, yeast sometimes participate in sexual reproduction, which is important because...
09:46
Expression of Recombinant Proteins in the Methylotrophic Yeast Pichia pastoris
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Cited by 59 •
2010
The protocol describes protein expression using the methylotrophic yeast Pichia pastoris. The preparation of electrocompetent yeast cells, transformation of the vector with the gene of interest into P. pastoris and yeast DNA purification are also performed. Western blot analysis and protein purification build the last steps in this protein expression protocol.
FAQs
Efficiency depends on how cells are prepared, the electrical conditions used for the pulse, the amount of DNA present, and the conditions provided afterward. These variables affect both molecular entry and cell recovery, so changing several at once can make results difficult to interpret. Systematic optimization helps produce transformation outcomes that are more reproducible across engineered yeast strains.
The electrical treatment temporarily disturbs the membrane, so cells need controlled recovery conditions after the pulse. Recovery gives cells an opportunity to restore membrane integrity and remain viable while the introduced genetic material becomes available for expression or maintenance. Inadequate recovery can reduce the number of usable transformed cells even when DNA entry has occurred.
A brief high-voltage pulse creates transient membrane pores rather than maintaining a permanent opening. These temporary disruptions allow DNA or another charged molecule to cross the membrane, after which the cells recover. The balance between electrical treatment and cellular recovery is therefore central to successful strain construction: the pulse must support molecular entry while preserving enough viable cells for downstream work.
A typical workflow begins with preparing the yeast cells and the DNA or other charged material, followed by exposing the cell suspension to a brief electrical pulse. The treated cells are then placed under controlled recovery conditions. Researchers evaluate the resulting cells for the intended genetic outcome, while adjusting preparation, pulse settings, DNA concentration, or recovery when reproducibility is poor.
Bioengineers can apply the method when they need to introduce genetic material into yeast for recombinant protein production, pathway engineering, or functional studies of metabolism. It is particularly relevant when constructing strains whose genetic changes must support a defined engineering objective. Because efficiency varies with experimental conditions, optimization becomes part of developing a reliable transformation workflow.
Successful treatment can produce yeast cells that express or maintain introduced genetic material, enabling researchers to examine engineered traits or metabolic functions. In synthetic biology, those outcomes support the construction of modified strains and the testing of designed pathways. The method therefore connects a physical delivery step with broader goals such as protein production, pathway engineering, and metabolic analysis.