Gfp Reporter Plasmid
Gfp Reporter Plasmid - Related Videos
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.
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.
08:16
Plasmid Purification
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2023
Plasmid purification is a technique used to isolate and purify plasmid DNA from genomic DNA, proteins, ribosomes, and the bacterial cell wall. A plasmid is a small, circular, double-stranded DNA that is used as a carrier of specific DNA molecules. When introduced into a host organism via transformation, a plasmid will be replicated, creating numerous copies of the DNA fragment under study. In this video, a step-by-step generalized procedure is described for how to perform plasmid purification.
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
The inserted regulatory sequence acts as the control point for GFP expression. When its associated promoter is active in a cell, host transcription machinery produces the corresponding RNA, and host translation machinery generates GFP. Fluorescence therefore reports activity linked to that regulatory element rather than serving as a universal readout of every cellular process. In neuroscience, this can connect signal patterns with molecular regulation.
The GFP chromophore emits green fluorescence only when exposed to suitable excitation light. Imaging conditions therefore determine whether expressed GFP becomes visible, while the signal itself reflects production of the fluorescent protein. This distinction helps researchers interpret live-cell images carefully, especially when comparing promoter activity or cellular changes across different observations over time.
Because the plasmid can couple GFP to a chosen regulatory sequence, fluorescence may indicate promoter-linked activity. When expression marks cells, the same signal can also outline neuronal shape. This allows researchers to examine molecular regulation and cellular morphology in one experimental system, while time-resolved imaging can extend the analysis to neuronal development or changing connectivity.
An informative signal depends on several linked conditions: the selected promoter or regulatory sequence must be active in relevant cells, the plasmid must be delivered, host transcription and translation must occur, and the GFP chromophore must be viewed with suitable excitation. Considering these factors helps distinguish absent regulatory activity from unsuccessful delivery or inadequate visualization.
After delivery into cells or tissue, the construct relies on host transcription and translation machinery to produce GFP. When the resulting protein is exposed to suitable excitation light, green fluorescence can be observed. Researchers can then image labeled neurons, examine their morphology, or monitor cellular activity linked to the selected regulatory sequence over time.
Live-cell imaging allows investigators to follow fluorescently labeled neurons as their appearance changes over time. Spatial patterns can help reveal neuronal morphology and connectivity, while promoter-linked expression can add information about molecular regulation. In cultured cells and experimental tissues, these observations connect dynamic structural changes with the cellular or regulatory context selected by the reporter design.
Following plasmid introduction, detectable GFP can provide a visible indication that the delivery or transfection experiment produced an expressed reporter in cells. Researchers can examine the fluorescent pattern in cultured cells or experimental tissues, then relate labeled cells to neuronal structure or regulatory activity. This makes the construct useful for connecting delivery outcomes with downstream imaging observations.