Photo-assisted Fabrication of Ribosome Display Microarray

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.