Reporter Protein Expression
Reporter Protein Expression - Related Videos
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.
03:15
Screening of Cyclic di-GMP Modulators Using Bacteria Expressing a Fluorescent Reporter Protein
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2025
Source: Rugjee, K. N., et al. Establishment of a High-throughput Setup for Screening Small Molecules That Modulate c-di-GMP Signaling in Pseudomonas aeruginosa. J. Vis. Exp. (2016).This video demonstrates the screening of cyclic di-GMP modulators using genetically modified bacteria that express green fluorescent protein (GFP) under the control of a c-di-GMP-sensitive promoter. Fluorescence is measured in a multi-well plate containing different test compounds. Compounds that inhibit c-di-GMP...
03:57
Preparation of Reporter-Expressing Cancer Cells for In Vivo Imaging
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2026
Source: Chen, H. et. al. Live Cell Imaging of the TGF-β/Smad3 Signaling Pathway In Vitro and In Vivo Using an Adenovirus Reporter System. J. Vis. Exp. (2018)This video demonstrates the preparation of luciferase-expressing breast cancer cells using recombinant adenoviruses for in vivo imaging.
11:40
Simplified Reverse Genetics Method to Recover Recombinant Rotaviruses Expressing Reporter Proteins
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Cited by 24 •
2020
Generation of recombinant rotaviruses from plasmid DNA provides an essential tool for the study of rotavirus replication and pathogenesis, and the development of rotavirus expression vectors and vaccines. Herein, we describe a simplified reverse genetics approach for generating recombinant rotaviruses, including strains expressing fluorescent reporter proteins.
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.
FAQs
Placing a reporter gene under a target promoter makes the measured signal an indicator of that regulatory sequence’s activity. Changes in signal can therefore be used to examine how strongly the promoter responds under different cellular conditions. This design is especially useful for comparing regulatory behavior during the optimization of engineered genetic circuits.
Fusing a reporter to a protein of interest connects the detectable signal to that protein rather than directly to promoter activity. The resulting measurement can provide information about protein localization, showing where the protein is present within the cell. This distinction helps bioengineers choose a design suited to studying regulation or intracellular distribution.
A reporter signal reflects the transcription and translation associated with the chosen genetic design, so its meaning depends on whether the reporter is controlled by a promoter or fused to a protein. Signal changes can indicate altered gene regulation or cellular state, but interpretation should remain tied to the specific reporter arrangement and measurement goal.
A typical workflow begins by selecting a target promoter or protein of interest, then positioning the reporter gene under that promoter or creating a protein fusion. Cells are evaluated for the resulting fluorescence or luminescence, and the signal is related to promoter activity, localization, or cellular-state changes. The findings can then guide genetic circuit design.
Both fluorescence and luminescence can serve as detectable outputs from a reporter system, but the appropriate choice depends on the measurement design and the information being sought. Researchers may select one signal type when evaluating promoter activity, protein localization, or cellular-state changes, provided the resulting readout matches the engineered construct and experimental objective.
Reporter measurements allow researchers to assess whether a regulatory sequence produces the intended activity and how strongly that activity changes. This feedback helps characterize and refine engineered genetic circuits rather than relying only on construct design. The same approach can also support evaluation of delivery strategies and analysis of biological processes in biotechnology.