Gfp Fusion Screening
Gfp Fusion Screening - Related Videos
10:30
The C. elegans Excretory Canal as a Model for Intracellular Lumen Morphogenesis and In Vivo Polarized Membrane Biogenesis in a Single Cell: labeling by GFP-fusions, RNAi Interaction Screen and Imaging
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Cited by 6 •
2017
The C. elegans excretory canal is a unique single-cell model for the visual in vivo analysis of de novo polarized membrane biogenesis. This protocol describes a combination of standard genetic/RNAi and imaging approaches, adaptable for the identification and characterization of molecules directing unicellular tubulogenesis, and apical membrane and lumen biogenesis.
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.
10:45
A Faster, High Resolution, mtPA-GFP-based Mitochondrial Fusion Assay Acquiring Kinetic Data of Multiple Cells in Parallel Using Confocal Microscopy
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Cited by 20 •
2012
Mitochondrial fusion was measured by tracking the equilibration of photoconverted matrix-targeted GFP across the mitochondrial network over time. Thus far, only one cell could be subjected to an hour long kinetic analysis at a time. We present a method that simultaneously measures multiple cells, thereby speeding up the data collection process.
02:41
Clonal Isolation and PCR Screening of GFP-Excised Recombinant Herpesvirus Retaining the Target Gene
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2026
Source: Tang, N., et al. Generating Recombinant Avian Herpesvirus Vectors with CRISPR/Cas9 Gene Editing. J. Vis. Exp. (2019)This video demonstrates the clonal isolation and PCR-based screening of recombinant herpesvirus following site-specific excision of a GFP marker. Fluorescence-activated cell sorting is used to isolate non-fluorescent infected cells, and junction PCR confirms retention of the target gene in GFP-negative clones.
02:45
Nuclear Fusion
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2020
The process of converting very light nuclei into heavier nuclei is also accompanied by the conversion of mass into large amounts of energy, a process called fusion. The principal source of energy in the sun is a net fusion reaction in which four hydrogen nuclei fuse and ultimately produce one helium nucleus and two positrons. A helium nucleus has a mass that is 0.7% less than that of four hydrogen nuclei; this lost mass is converted into energy during the fusion. This reaction produces about...
FAQs
The fusion places a visible reporter on the target protein, so cells that produce the recombinant construct can be identified through fluorescence after expression. Researchers can inspect individual cells by microscopy or quantify signal with fluorescence measurements, making fluorescence a practical readout for screening transformed populations and comparing production across constructs.
Because the fluorescent signal can be observed without staining or destroying the sample, researchers can examine protein behavior in living cells. This enables repeated assessment of expression, localization, and cellular distribution rather than relying only on a single endpoint. The approach therefore supports studies of trafficking and cellular organization while preserving cells for further observation.
Signal intensity can provide information about expression levels, while the position of fluorescence within a cell can indicate localization. In some cases, the observed pattern also helps assess solubility. These readouts act as complementary features of the expressed fusion, allowing researchers to evaluate production and intracellular behavior together rather than treating fluorescence as a simple presence-or-absence signal.
Researchers first create a genetic fusion between GFP and the target protein, then introduce or identify recombinant cells and allow expression. They next examine fluorescence by microscopy or fluorescence measurement. The resulting signal helps select successful cells or constructs and can guide subsequent evaluation of expression, localization, or solubility.
The method is useful when several recombinant constructs or expression conditions must be evaluated efficiently. Fluorescence provides a visible or measurable basis for selecting successful outcomes, while microscopy can add information about where the protein appears in cells. This makes screening relevant to both construct selection and optimization of protein production.
In biology, the approach supports studies of gene expression, protein trafficking, and cellular organization by linking a fluorescent signal to the target protein. It can also contribute to functional protein design and tracking behavior in living cells. The appropriate readout depends on whether the goal is production, localization, expression level, or intracellular behavior.