Team:Illinois/Antibody GPCR Fusion

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G protein-coupled receptors, or GPCRs, are transmembrane receptors sensing extracellular objects on the scale of small molecules to large proteins. Yeast cells are known to utilize two GPCRs signal transduction pathways, one to detect the presence of glucose and the other to initiate mating. We hope to engineer the well-characterized mating pathway to produce a colorimetric change in the cell upon detecting a novel molecule-- some water-bourne pathogen, specifically the beta subunit of cholera toxin.
G protein-coupled receptors, or GPCRs, are transmembrane receptors sensing extracellular objects on the scale of small molecules to large proteins. Yeast cells are known to utilize two GPCRs signal transduction pathways, one to detect the presence of glucose and the other to initiate mating. We hope to engineer the well-characterized mating pathway to produce a colorimetric change in the cell upon detecting a novel molecule-- some water-bourne pathogen, specifically the beta subunit of cholera toxin.
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==Specific Plans, Supplies, and Protocols==
In theory, this is how we will progress:
In theory, this is how we will progress:
*Create a fusion protein that links an antibody against cholera toxin to the Ste2 GPCR of S. cereviviae, the pheremone response GPCR.
*Create a fusion protein that links an antibody against cholera toxin to the Ste2 GPCR of S. cereviviae, the pheremone response GPCR.

Revision as of 18:54, 13 June 2008

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Contents

Core Team Members

Dave Luedtke, Bobak Hadidi

  • Add yourself!

Project Abstract

G protein-coupled receptors, or GPCRs, are transmembrane receptors sensing extracellular objects on the scale of small molecules to large proteins. Yeast cells are known to utilize two GPCRs signal transduction pathways, one to detect the presence of glucose and the other to initiate mating. We hope to engineer the well-characterized mating pathway to produce a colorimetric change in the cell upon detecting a novel molecule-- some water-bourne pathogen, specifically the beta subunit of cholera toxin.

Specific Plans, Supplies, and Protocols

In theory, this is how we will progress:

  • Create a fusion protein that links an antibody against cholera toxin to the Ste2 GPCR of S. cereviviae, the pheremone response GPCR.
    • 1)Find the sequences of the GPCR and the antibody
    • 2)Select site of fusion
    • 3)Have the gene sequenced
  • Express the antibody/GPCR fusion protein in yeast that lack the wild type receptor.
  • Measure the activation of the GPCR by the toxin (and by the natural pheremone) using a reporter gene.
  • Use site specific directed evolution to increase the effectiveness of the new GPCR.

Meetings

Fri. June 13th we will meet on the first floor of Grainger by the computers at 1:00pm

To Research

  • Cell Wall Issue
  • Antibody Sequence
  • Sources of yeast strains deficient in specific genes
  • Cholera Toxin or possible target protein
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