Gfp Reporter Construct
Gfp Reporter Construct - Related Videos
10:41
Using a GFP-tagged TMEM184A Construct for Confirmation of Heparin Receptor Identity
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Cited by 2 •
2017
A construct encoding TMEM184A with a GFP tag at the carboxy-terminus designed for eukaryotic expression, was employed in assays designed to confirm the identification of TMEM184A as a heparin receptor in vascular cells.
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.
06:19
In Vivo Imaging of Cx3cr1gfp/gfp Reporter Mice with Spectral-domain Optical Coherence Tomography and Scanning Laser Ophthalmoscopy
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Cited by 5 •
2017
This protocol describes how high-resolution imaging techniques such as spectral domain optical coherence tomography and scanning laser ophthalmoscopy can be utilized in small rodents, using an ophthalmic imaging platform system, to obtain information on retinal thickness and microglial cell distribution, respectively.
10:40
Characterization of Adipocyte-Derived Extracellular Vesicle Secretion Using a CD63-GFP Reporter Mouse Model In Vivo and In Vitro
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2025
The protocol uses an adipocyte-specific CD63-GFP reporter mouse line to visualize and quantify the secretion of adipocyte-derived extracellular vesicles (EVs) and demonstrate their uptake by progenitor cells, revealing a paracrine pathway within adipose tissue.
FAQs
Promoter selection determines which genetic program the construct reports. A promoter or regulatory sequence associated with pathogen expression can reveal when that program is active, whereas a sequence associated with immune-cell activation can mark responding cells. This makes the fluorescence pattern interpretable in relation to the selected regulatory element, rather than as an unspecified cellular signal.
Microscopy and flow cytometry answer different experimental questions. Fluorescence microscopy shows where GFP-positive activity occurs, helping visualize spatial patterns in cells or host-pathogen interactions. Flow cytometry measures fluorescence across many cells and can identify populations that are infected or responding. Using either readout supports quantitative analysis of infection dynamics.
Excitation is an essential part of GFP detection. The protein emits green light only after illumination with a suitable excitation wavelength, so the optical setup must provide that condition for fluorescence to be observed. In practice, this links the molecular activation event to a measurable signal that can be recorded by microscopy or flow cytometry.
Because GFP production follows activation of the linked regulatory sequence, changes in fluorescence can be used to follow changes in the selected genetic program. In infection studies, this supports tracking pathogen gene expression or host immune-cell activation over an experiment. The resulting signal can also help identify cells participating in the infection or response.
First, researchers connect the promoter or regulatory DNA sequence of interest to the GFP-encoding gene. They then examine whether the target program becomes active in the relevant experimental system and detect resulting fluorescence with microscopy or flow cytometry. This workflow connects regulatory activity with observable cellular or infection-related measurements.
In immunology and infection research, a GFP reporter construct can support several complementary purposes: tracking pathogen gene expression, monitoring immune-cell activation, observing host-pathogen interactions, or identifying infected and responding cells. The appropriate use depends on which promoter or regulatory sequence is linked to GFP and whether spatial visualization or population-level fluorescence is needed.
These reporters are useful when an experiment asks whether a regulatory mechanism or treatment changes a genetic response. Researchers can compare fluorescence-associated activity under the conditions being studied and use the readout to examine infection dynamics. Because the construct reports the linked program, interpretation should remain tied to that specific promoter or regulatory sequence.