Methods in Enzymology Vol.328 Applications of Chimeric Genes and Hybrid Proteins Part C

The accumulation of large amounts of genomic sequence data has prompted studies in protein biology on an unprecedented scale. Determining the functions of uncharacterized open reading frames (ORFs) from yeast and other organisms is a major challenge, and traditional biochemical and genetic approaches are struggling to keep up with sequence data. One strategy to help determine the functions is to identify protein–protein interactions using the two-hybrid system, a genetic assay that takes place within living yeast cells. An array format is developed in the chapter to increase the throughput of proteins that can be screened by the two-hybrid method. An array is a spatially ordered set of separated elements in which each element is a unique protein potentially available for interaction. This chapter describes methods for constructing and screening a protein array that contains most of the proteins present in S. cerevisiae. However, the array may represent any group of proteins: all the expressed proteins present in an organism a family of related proteins, or even a set of proteins or peptides, not found in nature.

• Series: Methods in Enzymology (Book 328)
• Hardcover: 666 pages
• Publisher: Academic Press;
  
• Language: English
• ISBN-10: 012182229X
• ISBN-13: 978-0121822293
• Product Dimensions: 9.2 x 6.3 x 1.3 inches

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