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Reporter Genes

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...

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Related Experiment Video

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Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli

Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli

Published on: January 6, 2015

Using superfolder green fluorescent protein for periplasmic protein localization studies.

Thuy Dinh¹, Thomas G Bernhardt

¹Department of Microbiology and Immunobiology, Harvard Medical School, Armenise Building, Room 302A, 200 Longwood Avenue, Boston, MA 02115, USA.

Journal of Bacteriology

|July 19, 2011

Summary

Superfolding green fluorescent protein (sfGFP) successfully tracks periplasmic protein export via Sec-mediated transport. This variant functions optimally when using the cotranslational pathway for protein translocation.

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Area of Science:

Cell Biology
Molecular Biology
Protein Biochemistry

Background:

Investigating subcellular protein localization is crucial for understanding cellular functions.
Traditional methods using green fluorescent protein (GFP) for periplasmic protein export studies are often unreliable.
Challenges exist in accurately visualizing protein transport pathways within the cell.

Purpose of the Study:

To develop a reliable method for studying the subcellular localization of periplasmic proteins.
To assess the utility of a superfolding variant of green fluorescent protein (sfGFP) in protein export studies.
To determine the optimal conditions for sfGFP fluorescence in relation to protein transport pathways.

Main Methods:

Utilized a superfolding variant of green fluorescent protein (sfGFP) as a reporter.
Investigated Sec-mediated protein transport pathways.
Employed cotranslational protein export mechanisms.
Assessed fluorescence of sfGFP following translocation across cellular membranes.

Main Results:

Demonstrated that sfGFP exhibits fluorescence after Sec-mediated transport.
Showcased that sfGFP is most effective when the cotranslational branch of the Sec pathway is utilized.
Overcame previous limitations associated with GFP export in localization studies.

Conclusions:

sfGFP provides a robust tool for monitoring periplasmic protein export.
The cotranslational pathway is a key determinant for successful sfGFP-based localization studies.
This advancement facilitates more accurate investigations into bacterial protein secretion and localization.