DARTs: A DNA-based in vitro polypeptide display technology

Proteomics
Paul de FigueiredoEugene W Nester

Abstract

Display technologies link proteins with the genes that encode them, providing a means of selecting proteins with desired properties through the process of directed evolution. Here, we describe DNA/protein attachment and recovery tools (DARTs), a novel polypeptide display technology that utilizes the Agrobacterium tumefaciens protein VirD2 to generate DNA-protein hybrid molecules. The resulting DNA-protein hybrids are small, robust, and are not expected to be subject to the synthesis and selection biases associated with viral- and cell-based display systems. We demonstrated that these DNA-protein hybrids could be used to display a variety of peptides that bind to appropriate antibodies for immunodetection and immunopurification. Further, the DNA components of the hybrid molecules can hybridize to complementary DNA molecules in solution or on a solid substrate. Because full-length VirD2 self-associated, we constructed a truncation that did not self-associate but still exhibited DNA linking activity and efficiently displayed peptides. Finally, we purified DNA-protein hybrids using their displayed peptide epitopes and amplified their DNA components by polymerase chain reaction. We suggest that the DART polypeptide display system wi...Continue Reading

Citations

May 8, 2013·Biologicals : Journal of the International Association of Biological Standardization·Nur Hidayah Hairul BaharaTheam Soon Lim
Jun 16, 2012·PloS One·Igor A KozlovMark S Chee
Sep 13, 2007·International Journal of Biological Sciences·Shingo UenoYuzuru Husimi
Aug 22, 2014·Nature Chemistry·Eugene J H Wee, Matt Trau
Oct 5, 2017·Nucleic Acids Research·Giulio Bernardinelli, Björn Högberg

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