Gerald Walter, Biorchard Limited, c/o Cornupia Capital Ltd, London, UK
Konrad Büssow, Max Planck Institute of Molecular Genetics, Berlin, Germany
Zoltan Konthur, Max Planck Institute of Molecular Genetics, Berlin, Germany
Angelika Lueking, Max Planck Institute of Molecular Genetics, Berlin, Germany
Jörn Glökler, Max Planck Institute of Molecular Genetics, Berlin, Germany
Ulrich Schneider, Scienion AG, Berlin, Germany
Published online: January 2006
DOI: 10.1038/npg.els.0005727
Array technology enables genome-wide screening for protein expression and interactions at high throughput. Protein microarrays are emerging as a major technology platform for functional genomics.
Keywords: protein; expression; interaction; microarray; arrayed library
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Figure 1. Genome-wide protein expression and interaction analysis using high-density filters and microarrays. cDNA expression libraries of tissue samples or antibody/peptide libraries are arrayed at high density using automated (robot) technology and are screened once for protein expression or antigen/ligand binding respectively. Positive clones (expressors or binders) are rearrayed into subset libraries. Microarrays are printed of (fusion) proteins, antibodies, peptides (or their expressing host cells), and are used for in vitro protein–X (e.g. –protein, –nucleic acid, –small molecule) or in vivo (yeast two-hybrid) interaction analysis.
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Figure 2. How the yeast two-hybrid system works. A truncated transcription factor is expressed in a yeast cell as two fusion proteins. One consists of a DNA-binding domain (DBD) fused to a protein X (bait). The other hybrid consists of a transcriptional activator domain (AD) fused to a protein Y (prey). If X and Y interact, the transcriptional activator is reconstructed and can activate transcription of a reporter system (e.g. HIS3, URA1, LacZ). Expression of HIS3 or URA1 enables the cell to grow on histidine- or uracil-free media, whereas expression of LacZ allows one to stain the cell. Therefore, interaction of two proteins generates an easily visible yeast colony.
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| Further Reading | |
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