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Related Concept Videos

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...

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Reporters of gene expression: autofluorescent proteins.

J E Rasko1

  • 1Centenary Institute of Cancer Medicine and Cell Biology, Sydney, Australia.

Current Protocols in Cytometry
|September 5, 2008
PubMed
Summary

This guide details Green Fluorescent Protein (GFP) transduction into cell lines, covering cell sorting, purification, and analysis. It offers troubleshooting and data for multiple GFP mutants, aiding researchers in successful gene expression studies.

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

  • Molecular Biology
  • Cell Biology
  • Biotechnology

Background:

  • Green Fluorescent Protein (GFP) is a vital reporter protein in biological research.
  • Efficiently introducing and analyzing GFP in cell lines is crucial for various experimental applications.

Purpose of the Study:

  • To provide a comprehensive protocol for the successful transduction of GFP into mammalian cell lines.
  • To offer detailed guidance on cell sorting, purification, and subsequent analysis of transduced cells.

Main Methods:

  • Detailed step-by-step instructions for GFP transduction.
  • Protocols for fluorescence-activated cell sorting (FACS) and cell purification.
  • Methods for analyzing GFP expression and validating transduced cells, including sample data.

Main Results:

  • Successful transduction of GFP into target cell lines was achieved following the provided protocol.
  • Purified and analyzed cell populations demonstrated high-quality GFP expression.
  • Troubleshooting strategies effectively addressed common challenges in the transduction process.

Conclusions:

  • The presented unit offers a robust and accessible method for GFP transduction and analysis.
  • This protocol facilitates reliable gene expression studies using GFP in various cell systems.
  • The inclusion of GFP mutant data and further reading enhances its utility for researchers.