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Genomic Analysis Utilizing the Yeast Two-Hybrid System

  • Ilya G. Serebriiskii
  • Garabet G. Toby
  • Russell L. FinleyJr.
  • Erica A. Golemis
Part of the Methods in Molecular Biology™ book series (MIMB, volume 175)

Abstract

As the completion of genome sequencing efforts leads to the definition of increasing numbers of genes, the need to reliably assign function to identified coding sequences becomes paramount. One means of gaining initial insight into the function of an undefined protein is to develop a map of other defined proteins with which it physically or functionally interacts. There are several approaches to assigning interacting protein groups. In suitable model organisms such as yeast, a traditional approach has been to create null mutations in the gene encoding the novel protein of interest, and to use suppressor analysis to identify genetically (functionally) interacting proteins. Alternatively, copurification of complexes of interest followed by use of mass spectrophotometry to assign identity of individual component proteins has been used to define interacting groups based on physical interactions. The genetic approaches offer speed and low cost; the physical approaches offer the certainty that copurified proteins physically function together on the protein level, rather than being connected via indirect regulatory pathways. A third approach, the yeast two-hybrid system, combines the advantages of working with yeast while targeting proteins that physically associate.

Keywords

Library Plasmid Library Screen Bait Protein Master Plate Bait Vector 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Humana Press Inc., Totowa, NJ 2001

Authors and Affiliations

  • Ilya G. Serebriiskii
    • 1
  • Garabet G. Toby
    • 1
  • Russell L. FinleyJr.
    • 2
  • Erica A. Golemis
    • 1
  1. 1.Division of Basic ScienceFox Chase Cancer CenterPhiladelphia
  2. 2.Center for Molecular Medicine and GeneticsWayne State University School of MedicineDetroit

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