Rapid isolation of novel FK506 binding proteins from multiple organisms using gDNA and cDNA T7 phage display

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Reverse chemical proteomics using T7 phage display is a powerful technique for identifying cellular receptors of biologically active small molecules. However, to date this method has generally been limited to cDNA libraries constructed from mRNA isolated from eukaryotes. In this paper, we describe the construction of the first prokaryotic T7 phage display libraries from randomly digested Pseudomonas stutzeri and Vibrio fischeri gDNA, as well as a plant cDNA library from Arabidopsis thaliana. We also describe the use of T7 phage display to identify novel proteins from environmental DNA samples using biotinylated FK506 as a model affinity probe.

LanguageEnglish
Pages6841-6850
Number of pages10
JournalBioorganic and Medicinal Chemistry
Volume17
Issue number19
DOIs
Publication statusPublished - 1 Oct 2009

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Tacrolimus Binding Proteins
Bacteriophage T7
Bacteriophages
Complementary DNA
Display devices
Gene Library
Pseudomonas stutzeri
Aliivibrio fischeri
Tacrolimus
Eukaryota
Arabidopsis
Proteomics
Libraries
Messenger RNA
Molecules
DNA
Proteins

Cite this

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abstract = "Reverse chemical proteomics using T7 phage display is a powerful technique for identifying cellular receptors of biologically active small molecules. However, to date this method has generally been limited to cDNA libraries constructed from mRNA isolated from eukaryotes. In this paper, we describe the construction of the first prokaryotic T7 phage display libraries from randomly digested Pseudomonas stutzeri and Vibrio fischeri gDNA, as well as a plant cDNA library from Arabidopsis thaliana. We also describe the use of T7 phage display to identify novel proteins from environmental DNA samples using biotinylated FK506 as a model affinity probe.",
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Rapid isolation of novel FK506 binding proteins from multiple organisms using gDNA and cDNA T7 phage display. / Piggott, Andrew M.; Kriegel, Alison M.; Willows, Robert D.; Karuso, Peter.

In: Bioorganic and Medicinal Chemistry, Vol. 17, No. 19, 01.10.2009, p. 6841-6850.

Research output: Contribution to journalArticleResearchpeer-review

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