Antibiotic Cassette
Antibiotic Cassette - Related Videos
03:36
Generating Gene Deletions in E. coli via Phage-Mediated Transfer of an Excisable Antibiotic Cassette
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2026
Source: Saragliadis, A., et al. Producing Gene Deletions in Escherichia coli by P1 Transduction with Excisable Antibiotic Resistance Cassettes. J. Vis. Exp. (2018)This video demonstrates the phage-mediated transfer of donor DNA carrying an excisable antibiotic resistance cassette into Escherichia coli. The integrated cassette replaces the target gene via homologous recombination, resulting in a stable gene deletion.
09:37
Screening Foodstuffs for Class 1 Integrons and Gene Cassettes
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Cited by 13 •
2015
This protocol describes the detection of class 1 integrons and their associated gene cassettes in foodstuffs.
08:13
Producing Gene Deletions in Escherichia coli by P1 Transduction with Excisable Antibiotic Resistance Cassettes
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Cited by 17 •
2018
Here we present a protocol for the use of pre-existing antibiotic resistance-cassette deletion constructs as a basis for making deletion mutants in other E. coli strains. Such deletion mutations can be mobilized and inserted into the corresponding locus of a recipient strain using P1 bacteriophage transduction.
11:03
Pouring and Running a Protein Gel by reusing Commercial Cassettes
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Cited by 2 •
2012
Our protocol demonstrates how to pour multiple protein gels at a time by recycling Invitrogen Nupage Novex minigel cassettes, and inexpensive materials purchased at a home improvement store. This economical and streamlined method includes a way to store the gels at 4°C for a few weeks. By re-using the plastic gel cassettes from commercially available gels, labs that run frequent protein gels can save significant costs and help the environment.
13:39
Antibiotic Susceptibility Testing: Epsilometer Tests to Determine MIC Values of Two Antibiotics and Evaluate Antibiotic Synergy
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2023
Source: Anna Bläckberg1, Rolf Lood1 1 Department of Clinical Sciences Lund, Division of Infection Medicine, Biomedical Center, Lund University, 221 00 Lund Sweden Knowledge of the interactions between antibiotics and bacteria is important in understanding how microbes evolve antibiotic resistance. In 1928, Alexander Fleming discovered penicillin, an antibiotic that exerts its antibacterial function by interfering with cell wall regeneration (1). Other antibiotics with diverse mechanisms of...
FAQs
Selection depends on coordinated expression of the resistance gene and its regulatory elements. The regulatory components determine whether the gene is expressed under the conditions used for growth, while the resistance gene supplies the trait that permits survival. This coupling makes antibiotic exposure a practical selection pressure for enriching cells that contain the engineered DNA.
The outcome depends on whether the introduced DNA is present and whether its cassette is expressed under the relevant growth conditions. Cells carrying an appropriately regulated resistance gene can survive antibiotic-containing medium, whereas nonmodified cells are eliminated. Surviving cells therefore provide a way to identify successful transformation events during genetic experiments.
Regulatory elements control expression of the resistance gene, so their compatibility with the growth conditions determines whether selection can operate. If the cassette is placed in a plasmid or genome and expressed appropriately, antibiotic exposure can distinguish cells containing the engineered DNA from nonmodified cells. Regulatory design is therefore central to reliable selection in genetic experiments.
A basic workflow begins by introducing DNA containing the cassette into cells, either through a plasmid or as part of a genomic modification. The cells are then grown on antibiotic-containing medium, where nonmodified cells are eliminated. Cells that survive can be identified as successful transformation events, supporting subsequent genetic analysis or strain development.
Antibiotic cassettes support several genetic strategies, including plasmid-based cloning, gene disruption, and transgene integration. In each case, the cassette provides a selectable feature that helps researchers identify cells associated with the intended DNA modification. This makes the approach useful for building engineered strains and experimental models in which modified genetic material must be distinguished from nonmodified material.
Researchers may remove or replace a cassette after it has served its selection purpose to reduce the genetic footprint of the engineered system. This refinement can improve the design of modified strains and experimental models by leaving less unnecessary selectable sequence behind. Cassette replacement also supports designs in which different genetic modifications require different selection arrangements.