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Related Experiment Videos

A genetic selection for isolating cDNAs encoding secreted proteins

K A Jacobs1, L A Collins-Racie, M Colbert

  • 1Genetics Institute, Inc., Cambridge, MA 02140, USA. kjacobs@genetics.com

Gene
|November 25, 1997
PubMed
Summary
This summary is machine-generated.

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This study introduces a rapid yeast-based genetic selection method for isolating cDNAs encoding secreted proteins. This technique efficiently identifies novel secreted proteins from large libraries without bioassays.

Area of Science:

  • Molecular Biology
  • Yeast Genetics
  • Protein Secretion

Background:

  • Identifying genes for secreted proteins is crucial for understanding cellular communication and function.
  • Existing methods for isolating secreted protein-encoding genes can be laborious and lack efficiency.
  • A robust and scalable method is needed to discover novel secreted proteins.

Purpose of the Study:

  • To develop and validate a novel, rapid genetic selection technique for isolating cDNAs encoding secreted proteins.
  • To demonstrate the utility of this method for identifying diverse secreted proteins from human cells.
  • To enable the discovery of novel secreted proteins, including chemokines and G-protein-coupled receptors.

Main Methods:

  • Utilized a Saccharomyces cerevisiae strain deficient in endogenous invertase.

Related Experiment Videos

  • Employed a cDNA cloning vector with a modified invertase gene lacking its leader sequence.
  • Applied a microbial growth selection strategy based on sucrose or raffinose utilization.

    • Main Results:

    • Successfully isolated large numbers of cDNAs encoding secreted proteins from a human peripheral blood mononuclear cell (PBMC) library.
    • Identified genes encoding diverse proteins, including cytokines, transmembrane proteins, and organelle-associated proteins.
    • Discovered novel secreted proteins, such as a chemokine and a G-protein-coupled receptor.

    Conclusions:

    • The described yeast-based genetic selection is a powerful and efficient tool for discovering novel secreted proteins.
    • The technique is broadly applicable for isolating secreted protein-encoding genes from various eukaryotic and prokaryotic organisms.
    • This method significantly advances the field of secreted protein discovery by bypassing the need for specific bioassays.