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Related Concept Videos

Tagging and Fusion Proteins01:24

Tagging and Fusion Proteins

Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...
Reporter Genes02:11

Reporter Genes

Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
Commonly used reporter...
FISH - Fluorescent In-situ Hybridization02:07

FISH - Fluorescent In-situ Hybridization

Fluorescence in situ hybridization, or FISH, was developed in the early 1980s and has quickly become one of the most widely used techniques in cytogenetics. Labeled probes are used to bind complementary DNA or RNA sequences on a chromosome or in a region within a cell. Earlier, the probes could only be obtained by cloning or reverse transcription of a DNA template. Currently, the probe oligonucleotides can be synthesized synthetically. Additionally, with the advancement of optical techniques,...

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Related Experiment Video

Updated: Jun 7, 2026

Imaging Subcellular Structures in the Living Zebrafish Embryo
11:19

Imaging Subcellular Structures in the Living Zebrafish Embryo

Published on: April 2, 2016

Constructing and expressing fluorescent protein fusions.

David L Spector, Robert D Goldman

    Cold Spring Harbor Protocols
    |November 3, 2010
    PubMed
    Summary
    This summary is machine-generated.

    Fluorescent protein fusions (FPFs) enable cell and tissue research, but careful design is crucial. Ensuring correct expression, localization, and function of FPFs is vital for accurate experimental results.

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    Last Updated: Jun 7, 2026

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    Bimolecular Fluorescence Complementation
    08:54

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    Published on: April 15, 2011

    Co-expression of Multiple Chimeric Fluorescent Fusion Proteins in an Efficient Way in Plants
    09:45

    Co-expression of Multiple Chimeric Fluorescent Fusion Proteins in an Efficient Way in Plants

    Published on: July 1, 2018

    Area of Science:

    • Cell Biology
    • Molecular Biology
    • Biochemistry

    Background:

    • Fluorescent protein fusions (FPFs) are widely used tools in biological research.
    • Their application spans individual cells to specific tissues within organisms.
    • Potential challenges include ensuring the fusion protein's expression level, integrity, localization, and normal cellular behavior.

    Purpose of the Study:

    • To discuss the critical considerations for successfully using FPFs in biological research.
    • To provide guidance on achieving accurate experimental outcomes with FPFs.
    • To offer examples for designing and implementing FPF experiments.

    Main Methods:

    • Review of best practices for FPF experimental design.
    • Consideration of the impact of the fluorescent protein (FP) tag's molecular mass (27 kDa).
    • Strategies for validating FPF expression, localization, and function.

    Main Results:

    • FPFs require careful validation to ensure they mimic endogenous protein behavior.
    • The 27 kDa mass of the FP tag can influence protein function and localization.
    • Successful FPF use depends on meticulous experimental design and verification.

    Conclusions:

    • Investigators must exercise caution when designing and using FPFs.
    • Proper validation is essential to mitigate potential artifacts introduced by the FP tag.
    • Adherence to best practices ensures the reliability of FPF-based research findings.