From Electronic Sequence to Purified Protein Using Automated Gene Synthesis and Transcription/Translation.

From Electronic Sequence to Purified Protein Using Automated Gene Synthesis and Transcription/Translation.

Chiocchini, Claudia;Vattem, Krishna;Liss, Michael;Ludewig, Lisa;Reusch, Tobias;Rastogi, Ischwaku;Webb, Brian;Trefzer, Axel;
acs synthetic biology 2020 Vol. 9 pp. 1714-1724
147
chiocchini2020fromacs

Abstract

gene synthesis is the state-of-the-art method used to obtain genetic material adapted to the requirements of the host organism and a cornerstone for modern synthetic biology. Yet, little progress has been made regarding downstream processes of protein production from synthetic genetic material. The production of recombinant proteins traditionally requires extensive preparatory work including gene amplification, cloning, sequencing, transformation or transfection of the expression host, cultivation of living cells, and purification of the overexpressed protein. In this work we describe a fast and automated workflow for cell-free production of proteins starting from an electronic protein sequence or accession number. PRESTO (protein expression starting from oligonucleotides) seamlessly combines a tailored sequence optimization with the assembly of short oligonucleotides into synthetic linear DNA expression cassettes, mammalian transcription/translation, and protein purification thereof. Integrated on a small liquid handling system it provides a hands-free high throughput source for functional synthetic proteins within 1 day.

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ID: 109639
Ref Key: chiocchini2020fromacs
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0x95644003c57E6F55A65596E3D9Eac6813e3566dA
Article ID:
109639
Unique Identifier:
10.1021/acssynbio.0c00060
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