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De novo design and synthesis of a 30-cistron translation-factor module
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology, Structure and Molecular Biology. MIT, Dept Biol Engn, Cambridge, MA 02139 USA..
Vanderbilt Univ, Med Ctr, Dept Pharmacol, Nashville, TN 37232 USA.;Vanderbilt Univ, Med Ctr, Ctr Quantitat Sci, Nashville, TN 37232 USA..
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology, Structure and Molecular Biology.
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology, Structure and Molecular Biology.
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2017 (English)In: Nucleic Acids Research, ISSN 0305-1048, E-ISSN 1362-4962, Vol. 45, no 18, p. 10895-10905Article in journal (Refereed) Published
Abstract [en]

Two of the many goals of synthetic biology are synthesizing large biochemical systems and simplifying their assembly. While several genes have been assembled together by modular idempotent cloning, it is unclear if such simplified strategies scale to very large constructs for expression and purification of whole pathways. Here we synthesize from oligodeoxyribonucleotides a completely de-novo-designed, 58-kb multigene DNA. This BioBrick plasmid insert encodes 30 of the 31 translation factors of the PURE translation system, each His-tagged and in separate transcription cistrons. Dividing the insert between three high-copy expression plasmids enables the bulk purification of the aminoacyl-tRNA synthetases and translation factors necessary for affordable, scalable reconstitution of an in vitro transcription and translation system, PURE 3.0.

Place, publisher, year, edition, pages
OXFORD UNIV PRESS , 2017. Vol. 45, no 18, p. 10895-10905
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Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
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URN: urn:nbn:se:uu:diva-340137DOI: 10.1093/nar/gkx753ISI: 000413107400046PubMedID: 28977654OAI: oai:DiVA.org:uu-340137DiVA, id: diva2:1178429
Funder
NIH (National Institute of Health), R01-AI0727453Swedish Research Council, NT 2011-5787Swedish Research Council, NT 2016-1Swedish Research Council, 2016-1
Available from: 2018-01-29 Created: 2018-01-29 Last updated: 2018-01-29Bibliographically approved

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