Gfp Reporter Expression
Gfp Reporter Expression - Related Videos
03:38
Measuring Mycobacterium smegmatis Mistranslation Rates Using a GFP/Luciferase Reporter System
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2025
Source: Chen, Y., et al., Measurement of Specific Mycobacterial Mistranslation Rates with Gain-of-function Reporter Systems. J. Vis. Exp. (2019)This video demonstrates a method to quantify mistranslation in Mycobacterium smegmatis using an inducible GFP and secreted mutant luciferase reporter system. It shows how luminescence resulting from mistranslation is measured and compared to GFP fluorescence to evaluate the impact of a test molecule on translational accuracy.
09:22
Rapid Synthesis and Screening of Chemically Activated Transcription Factors with GFP-based Reporters
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Cited by 11 •
2013
This protocol describes an experimental procedure for the rapid construction of artificial transcription factors (ATFs) with cognate GFP reporters and quantification of the ATFs ability to stimulate GFP expression via flow cytometry.
10:58
Assessment of GFP Expression and Viability Using the Tali Image-Based Cytometer
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Cited by 14 •
2011
This protocol describes how to perform cell viability and fluorescence expression assays using the Tali Image-Based Cytometer.
05:28
Targeted Expression of GFP in the Hair Follicle Using Ex Vivo Viral Transduction
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Cited by 1 •
2008
The hair follicle is a highly complex appendage of the skin containing a multiplicity of cell types. The follicle undergoes constant cycling through the life of the organism including growth and resorption with growth dependent on specific stem cells. The targeting of the follicle by genes and stem cells to change its properties, in particular, the nature of the hair shaft is, discussed. Hair follicle delivery systems are described, such as liposomes and viral vectors for gene therapy. The...
07:30
Electrophysiological Characterization of GFP-Expressing Cell Populations in the Intact Retina
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Cited by 5 •
2011
This article depicts the recording of individual cells from fluorescently tagged neuronal populations in the intact mouse retina. By using two-photon infrared excitation transgenetically labeled cells were targeted for patch-clamp recording to study their light responses, receptive field properties, and morphology.
FAQs
The promoter or another regulatory element determines when and where the GFP coding sequence is transcribed. Because GFP production follows that regulatory activity, changes in fluorescence can reveal differences in gene expression patterns across cells, tissues, or developmental stages. Selecting the linked regulatory sequence therefore determines which biological activity the reporter can visualize.
A visible GFP signal depends on several linked steps: the reporter sequence must be transcribed, the resulting RNA must be translated into protein, and the protein must form its fluorescent chromophore. After excitation, that chromophore emits green light. Consequently, fluorescence reflects the successful production and maturation of the reporter rather than transcription alone.
The spatial distribution and timing of fluorescence can show where a gene or pathway is active and how that activity changes during development or across tissues. When the reporter is associated with a protein or cellular process, fluorescence microscopy can also provide information about protein localization and dynamic cellular behavior in living material.
Researchers place the GFP coding sequence under the control of a promoter or other regulatory element selected for the biological activity of interest. They then examine the resulting cells, tissues, or organisms with fluorescence microscopy, using excitation to produce the green signal. Comparing fluorescence across locations or times helps interpret the linked gene or pathway activity.
Fluorescence microscopy supplies the excitation needed to make the GFP chromophore emit detectable green light and allows that signal to be observed within biological samples. This makes it possible to examine patterns in cells, tissues, or whole organisms while preserving a noninvasive view of activity over time, supporting real-time observation of cellular and developmental changes.
The approach is useful when researchers need to monitor gene expression patterns, pathway activity, protein localization, or developmental changes in living material. Its applications span molecular biology, genetics, and cell biology because the same visual signal can connect regulatory activity with its location and timing without requiring invasive observation of every change.