Egfp Gene
Egfp Gene - Related Videos
13:34
Gene Trapping Using Gal4 in Zebrafish
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Cited by 19 •
2013
This protocol describes the method of gene trap insertional mutagenesis using Gal4-VP16 as the primary reporter and GFP/RFP as secondary reporters in zebrafish. Approximately one in ten high-expressing F0 fish yield gene trap progeny co-expressing GFP and RFP. The screening procedure can be readily scaled to adapt to the size of the laboratory performing the insertional mutagenesis screen.
11:43
Easy Measurement of Diffusion Coefficients of EGFP-tagged Plasma Membrane Proteins Using k-Space Image Correlation Spectroscopy
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Cited by 37 •
2014
This paper provides a step by step guide to the fluctuation analysis technique k-Space Image Correlation Spectroscopy (kICS) for measuring diffusion coefficients of fluorescently labeled plasma membrane proteins in live mammalian cells.
10:03
Development of Leishmania Species Strains with Constitutive Expression of eGFP
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2023
Here, we describe the methodology used for generating L. panamensis and L. donovani strains expressing the gene for eGFP as a stable integrated transgene using the pLEXSY system. Transfected parasites were cloned by limiting dilution, and clones with the highest fluorescence intensity in both species were selected for further use in drug screening assays.
05:44
Gene-gun Transfection of Hippocampal Neurons
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Cited by 2 •
2006
10:20
Studying Membrane Protein Trafficking in Drosophila Photoreceptor Cells Using eGFP-Tagged Proteins
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Cited by 1 •
2022
Here, non-invasive methods are described for localization of photoreceptor membrane proteins and assessment of retinal degeneration in the Drosophila compound eye using eGFP fluorescence.
FAQs
After translation, the EGFP protein undergoes an autocatalytic modification involving selected amino acids that creates its internal chromophore. Because this light-producing structure forms within the protein itself, researchers do not need to supply an external substrate to generate fluorescence. Excitation with suitable light then makes the protein emit green fluorescence for direct observation in living cells.
These designs answer different genetic questions. Linking EGFP to a selected regulatory sequence makes fluorescence report activity controlled by that sequence, such as promoter activity. Fusing EGFP to a gene of interest instead allows researchers to follow the associated protein's cellular localization. The first emphasizes regulation of expression, whereas the second emphasizes where a protein is found.
EGFP converts genetic activity into an observable and measurable fluorescent signal. When its expression is controlled by selected regulatory sequences or linked to a gene of interest, changes in fluorescence can be examined alongside cellular location or behavior. This connection helps researchers interpret how genetic information relates to transcription, protein targeting, and developmental changes in living cells.
A typical design begins by choosing whether EGFP should report regulatory activity or accompany a protein of interest. Researchers then place the gene under the selected regulatory sequence or create a fusion with the target gene. Following introduction into cells, suitable illumination reveals fluorescence, allowing the selected genetic activity or protein localization to be examined.
EGFP can provide a visible readout of whether cells received and expressed introduced genetic material. After transfection, researchers examine green fluorescence in the cell population and use its presence as an indicator of successful expression. This application helps distinguish cells showing reporter activity from those without a detectable signal, supporting evaluation of the genetic delivery step.
When EGFP expression is linked to selected genetic control elements, fluorescence can be followed across cells and developmental stages. Researchers can track which cells retain or display the reporter, helping examine lineage patterns and inheritance. In developmental studies, these observations provide a visual way to relate changing genetic activity to the emergence of biological structures or behaviors.