Reporter Protein Expression
Reporter Protein Expression - Related Videos
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...
08:46
Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli
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Cited by 33 •
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.
08:07
Profiling of Surface Protein Epitopes on Viral Particles by Multiplex Dual-Reporter Strategy
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2024
Here, we describe a newly developed multiplex fluorescent immunoassay that uses a dual-reporter flow cytometric system to concurrently detect two unique spike protein epitopes on intact severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) viral particles that had been captured by angiotensin-converting enzyme-2-coupled magnetic microspheres.
11:40
Simplified Reverse Genetics Method to Recover Recombinant Rotaviruses Expressing Reporter Proteins
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Cited by 20 •
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.
10:48
Vaccinia Reporter Viruses for Quantifying Viral Function at All Stages of Gene Expression
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Cited by 4 •
2014
We describe the usage of a fluorescent reporter vaccinia virus that enables real-time measurement of viral infectivity and gene expression through the stage-specific expression of spectrally distinct reporter fluorophores. We detail a plate-based method for accurately identifying the stage at which virus replication is affected in response to small molecule inhibition.
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.