Photo-assisted Fabrication of Ribosome Display Microarray

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  • Kumal Subhashini Raj
    Department of Bioengineering, School of Engineering, The University of Tokyo
  • Kobayashi Ryo
    Department of Bioengineering, School of Engineering, The University of Tokyo
  • Ueno Shingo
    Department of Bioengineering, School of Engineering, The University of Tokyo Core Research of Evolutional Science and Technology, Japan Science and Technology Agency
  • Ichiki Takanori
    Department of Bioengineering, School of Engineering, The University of Tokyo Core Research of Evolutional Science and Technology, Japan Science and Technology Agency

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Ribosome display has been widely used for in vitro protein evolution, and the formation of a ternary complex, mRNA-ribosome-protein, is central to this technology as it displays the genotype-phenotype linkage. In conventional ribosome display methods, DNA with a native sequence containing stop codons cannot be used as a starting material as the ribosome in the ternary complex is primarily stalled by the removal of stop codons. Here, a new method for ribosome display is reported. This new method enables the use of DNA containing a stop codon as a starting material and presents the ribosome display on a microarray format with the assistance of a photo-crosslinker. This method is expected to be more advantageous than the conventional method as it allows the use of DNA with a native sequence without any modification, and since it is presented on a microarray format, it enables immediate functional identification and comparison between different proteins without any downstream processing.

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