Microintaglio printing of biomolecules and its application to in situ production of messenger ribonucleic acid display microarray

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A new molecular printing technology for a simple and robust patterning of biomolecules was developed and applied to producing a genotype-linked protein microarray. A microarray of messenger ribonucleic acids (mRNAs) encoding the green fluorescent protein was patterned by microintaglio printing using a micromold comprising an array of uniformly arranged 5-µm-diameter holes at a density of 10,000 per mm2. Furthermore, one-step conversion from an mRNA microarray into an mRNA-protein fusion microarray was performed by simultaneous cell-free protein synthesis and fusion reaction using a puromycin-labeled oligonucleotide linker. The set of developed technologies provides a powerful means of in vitro protein evolution.

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