Luciferase Protein Expression
Luciferase Protein Expression - Related Videos
08:09
High-throughput Titration of Luciferase-expressing Recombinant Viruses
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Cited by 14 •
2014
This article presents a high-throughput luciferase expression-based method of titrating various RNA and DNA viruses using automated and manual liquid handlers.
04:02
Split Luciferase Complementation Assay to Identify Specific Protein-Protein Interactions
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2025
This video demonstrates the split luciferase complementation assay to detect protein-protein interactions. In this assay, the proteins of interest are tagged to small and large fragments of the luciferase enzyme. When the proteins interact, the large and small fragments combine to form an active enzyme complex, which in the presence of a specific substrate, releases bright luminescence that can be measured.
03:23
Measurement of the In Vivo Burden of Bacteria Expressing Luciferase in an Insect Larva
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2025
This video showcases the in vivo bacterial burden measurement using Bacillus Calmette-Guérin, BCG, lux-infected insect larvae. The process involves larval homogenization, treatment with a luciferase substrate, and bioluminescence measurement to determine relative light units that correlate with bacterial burden.
10:38
Monitoring Cell-autonomous Circadian Clock Rhythms of Gene Expression Using Luciferase Bioluminescence Reporters
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Cited by 78 •
2012
Circadian clocks function within individual cells, i.e., they are cell-autonomous. Here, we describe methods for generating cell-autonomous clock models using non-invasive, luciferase-based real-time bioluminescence technology. Reporter cells provide tractable, functional model systems for studying circadian biology.
08:46
Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli
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Cited by 34 •
2015
A streamlined approach to screening for the expression of recombinant membrane proteins in Escherichia coli based on fusion to green fluorescent protein is presented.
FAQs
The promoter or other regulatory element determines when and how strongly the luciferase coding sequence is expressed in a cell. Changing that control region lets researchers test promoter function or enhancer regulation rather than merely detect protein production. Differences in emitted signal can reveal how specific regulatory sequences alter gene activity under the tested cellular conditions.
Signal intensity can serve as a readout at more than one regulatory level. A change may reflect altered transcription, altered translation, or activation of a cellular pathway that affects reporter output. Consequently, the light measurement is useful for comparing experimental conditions, but its meaning depends on which regulatory process the reporter construct was designed to monitor.
Luciferase catalyzes oxidation of the luciferin substrate, and that reaction generates bioluminescence that can be measured. The resulting light provides a sensitive, quantitative output linked to reporter activity. This chemical step converts molecular changes in gene regulation or pathway activity into a signal suitable for comparing samples or experimental treatments.
Researchers first place a luciferase coding sequence under a selected promoter or regulatory element and introduce that reporter into living cells. After the cells express the enzyme, the luciferin substrate is used in the assay and emitted light is measured. The resulting signal is then interpreted in relation to the regulatory question or experimental condition being tested.
It is useful for evaluating promoter function, enhancer regulation, or cellular responses to an experimental effect. The same reporter strategy can also provide information about transfection efficiency, helping assess how effectively genetic material reached cells. These applications make the assay a flexible readout for testing regulatory elements and comparing responses in living-cell experiments.
In genetics, the method links a chosen regulatory sequence to a measurable light output. Researchers can use that relationship to dissect how promoters and enhancers influence gene activity and to evaluate changes caused by experimental conditions. Because the output is quantitative, comparisons can help distinguish stronger or weaker regulatory effects within the assay.