Lux Gene Cassette
Lux Gene Cassette - Related Videos
08:10
A Functional Whole Blood Assay to Measure Viability of Mycobacteria, using Reporter-Gene Tagged BCG or M.Tb (BCG lux/M.Tb lux)
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Cited by 19 •
2011
We describe an alternative approach to the enumeration of mycobacteria in vitro, which uses reporter-gene tagged mycobacteria instead of colony-forming units (CFU). “Survival” of organisms as well as host response-markers are measured simultaneously, providing a low-cost, versatile and functional system for studies of host/pathogen interactions in the context of tuberculosis.
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.
04:06
Preparation and Quantification of Mycobacterium bovis BCG lux Inoculum for Infection
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2025
This video demonstrates the preparation and quantification of Mycobacterium bovis Bacillus Calmette-Guérin lux inoculum using bioluminescence assay and the enumeration of colony-forming units. The prepared inoculum can be used for infection studies.
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.
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.
FAQs
The two gene groups provide complementary functions. luxAB encodes luciferase, the enzyme that produces light, while luxCDE supports synthesis and recycling of the long-chain aldehyde required for the reaction. This arrangement helps bacterial cells maintain the substrate needed for repeated light production, making the signal suitable for monitoring ongoing biological activity rather than a single isolated event.
Light production requires luciferase, reduced flavin mononucleotide, oxygen, and a long-chain aldehyde. Luciferase uses these components in the bioluminescent reaction, so the signal reflects whether the cassette's enzymatic and substrate-producing functions can operate within the bacterial cell. In infection studies, this chemistry provides the basis for detecting microbial activity through emitted light.
Repeated measurements allow researchers to follow the same biological process over time without relying only on a single endpoint. Changes in light output can be related to pathogen activity during infection, colonization, or dissemination and then compared with evolving host immune responses. This temporal connection helps reveal how microbial behavior changes during disease or intervention.
Researchers introduce the cassette into a microbial pathogen and then measure the light produced during the experiment. Serial measurements can track where and when the pathogen remains active, colonizes, or disseminates. The resulting signal provides a noninvasive way to follow infection dynamics over time and to examine how those dynamics correspond with host responses.
Lux reporters can support monitoring of pathogen colonization, dissemination, and general infection activity. Tracking signal repeatedly helps show whether microbial behavior changes as an infection progresses, rather than providing information only at one selected time. In immunology and infection studies, this makes the reporter useful for relating pathogen distribution or activity to the host immune response.
Light output can be monitored before and after an antimicrobial or immune-based intervention to assess changes in pathogen activity over time. A changing signal provides a readout of how the microbial infection responds while host immunity or treatment is being studied. This approach can connect intervention-related effects with the underlying course of infection and immune response.