cDNA Libraries pp 199-210 | Cite as

Construction of Yeast Surface-Displayed cDNA Libraries

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 729)

Abstract

Using yeast display, heterologous protein fragments can be efficiently displayed at high copy levels on the Saccharomyces cerevisiae cell wall. Yeast display can be used to screen large expressed protein libraries for proteins or protein fragments with specific binding properties. Recently, yeast surface-displayed cDNA libraries have been constructed and used to identify proteins that bind to various target molecules such as peptides, small molecules, and antibodies. Because yeast protein expression pathways are similar to those found in mammalian cells, human protein fragments displayed on the yeast cell wall are likely to be properly folded and functional. Coupled with fluorescence-activated cell sorting, yeast surface-displayed cDNA libraries potentially allow the selection of protein fragments or domains with affinity for any soluble molecule that can be fluorescently detected. In this report, we describe protocols for the construction and validation of yeast surface-displayed cDNA libraries using preexisting yeast two-hybrid cDNA libraries as a starting point.

Key words

Yeast Surface cDNA display Library 

Notes

Acknowledgments

The work is supported by grants from the National Institute of Health (R01 CA118919, R01 CA129491, R21 CA137429, and R21 CA135586).

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.UCSF Comprehensive Cancer CenterUniversity of California at San FranciscoSan FranciscoUSA
  2. 2.Department of Anesthesia, UCSF Comprehensive Cancer CenterUniversity of California at San FranciscoSan FranciscoUSA

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