Egfp Expressing Cells
Egfp Expressing Cells - Related Videos
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.
10:34
Using an Automated Cell Counter to Simplify Gene Expression Studies: siRNA Knockdown of IL-4 Dependent Gene Expression in Namalwa Cells
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Cited by 2 •
2010
This procedure describes a quick and easy workflow to introduce siRNA into difficult to transfect cell lines and follow gene expression by real-time PCR. Use of an automated cell counter, multi-well electroporation plate, and automated electrophoresis station provide quick and reliable results without the need for expensive robotic handling.
12:09
Transient Expression and Cellular Localization of Recombinant Proteins in Cultured Insect Cells
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Cited by 6 •
2017
Nonlytic insect cell expression systems are underutilized for production, cellular trafficking/localization, and recombinant protein functional analysis. Here, we describe methods to generate expression vectors and subsequent transient protein expression in commercially available lepidopteran cell lines. The co-localization of Bemisia tabaci aquaporins with subcellular fluorescent marker proteins is also presented.
10:54
Reliable Identification of Living Dopaminergic Neurons in Midbrain Cultures Using RNA Sequencing and TH-promoter-driven eGFP Expression
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Cited by 9 •
2017
In Parkinson's Disease (PD), Substantia Nigra (SNc) dopaminergic neurons degenerate, leading to motor dysfunction. Here we report a protocol for culturing ventral midbrain neurons from a mouse expressing eGFP driven by a Tyrosine Hydroxylase (TH) promoter sequence, harvesting individual fluorescent neurons from the cultures, and measuring their transcriptome using RNA-seq.
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, EGFP must fold into the appropriate structure before forming its light-absorbing chromophore. This chromophore gives the protein its ability to emit green fluorescence after excitation with blue or ultraviolet light. Consequently, visible signal depends not only on production of the protein, but also on successful folding and chromophore formation within the living cell.
The signal connects several stages of gene activity: DNA containing the EGFP sequence is transcribed into messenger RNA, and that RNA is translated into EGFP protein. Fluorescence therefore provides a visible readout of cells that have processed the introduced genetic information through these steps. This makes the marker useful for observing gene expression in living biological systems.
Fluorescence microscopy can show where EGFP-containing cells or labeled proteins appear within a biological sample, while the presence of signal indicates that expression has occurred. This combination supports studies of cell location, movement, and protein localization rather than limiting analysis to whether a construct entered the cells. The resulting observations can be collected over time in living systems.
Researchers first introduce an EGFP-containing DNA construct into cells. The cells then transcribe the inserted gene, translate its messenger RNA, and produce EGFP protein that folds and forms its chromophore. Once fluorescence develops, researchers can examine the cells with fluorescence microscopy or analyze them with flow cytometry, depending on whether spatial or population-level information is needed.
Fluorescence microscopy is especially useful when the research question depends on visual location. It can reveal where labeled cells are positioned, whether they move, and where an EGFP-associated protein appears within the cell. Because observations can be made in living cells, this approach supports real-time examination of cellular behavior and protein localization in biological studies.
Flow cytometry provides a complementary way to analyze EGFP fluorescence across cells rather than focusing primarily on their spatial arrangement. It is useful for assessing transfection, meaning the introduction of the DNA construct into cells, and for examining fluorescence within a cell population. Together with microscopy, it broadens analysis from visual tracking to cell-based measurement.