Use of Rolling-Circle Amplification for Large-Scale Yeast Two-Hybrid Analyses

  • Xiaodong Ding
  • Yan Zhang
  • Wen-Yuan Song
Part of the Methods in Molecular Biology book series (MIMB, volume 354)

Abstract

Detection of protein–protein interactions on a large-scale has become a major focus of functional genomics after the completion of genome sequencing. The information generated from these studies not only assembles proteins into signaling networks, but also reveals potential functions of uncharacterized proteins when their interacting partners have known functions. We have developed a rolling circle amplification-based yeast two-hybrid scheme that allows one to test reproducibility and specificity of the interactions on a large scale. Using this scheme, technical false-positives from yeast two-hybrid analyses can be efficiently minimized.

Key Words

Protein—protein interactions yeast two-hybrid rolling-circle amplification high throughput plasmids 

Notes

Acknowledgments

We thank Dr. M. Vidal for providing the yeast strain MaV203, Dr. P. C. Ronald for the precursors of the pXDGATcy86 and pXDGATU86 vectors, and Lisa Nodzon for critical reading of the manuscript. This research was supported by the Florida Agricultural Experiment Station and a grant from the National Science Foundation Plant Genome Research to W-Y S. This work was approved for publication as Journal Series No. R-10865.

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Copyright information

© Humana Press Inc. 2007

Authors and Affiliations

  • Xiaodong Ding
    • 1
  • Yan Zhang
    • 2
  • Wen-Yuan Song
    • 1
  1. 1.Department of Plant PathologyUniversity of FloridaGainesville
  2. 2.University of FloridaGainesville

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