An efficient method for the construction of artificial, concatemeric DNA, RNA and proteins with genetically programmed functions, using a novel, vector-enzymatic DNA fragment amplification-expression technology.

Autor:	Skowron PM; Department of Molecular Biotechnology, Faculty of Chemistry, University of Gdansk, Gdansk 80-308, Poland.; BioVentures Institute Ltd., Poznan 60-141, Poland., Krawczun N; Department of Molecular Biotechnology, Faculty of Chemistry, University of Gdansk, Gdansk 80-308, Poland.; BioVentures Institute Ltd., Poznan 60-141, Poland., Żebrowska J; Department of Molecular Biotechnology, Faculty of Chemistry, University of Gdansk, Gdansk 80-308, Poland.; BioVentures Institute Ltd., Poznan 60-141, Poland., Krefft D; Department of Molecular Biotechnology, Faculty of Chemistry, University of Gdansk, Gdansk 80-308, Poland.; BioVentures Institute Ltd., Poznan 60-141, Poland., Żołnierkiewicz O; Department of Molecular Biotechnology, Faculty of Chemistry, University of Gdansk, Gdansk 80-308, Poland., Bielawa M; BioVentures Institute Ltd., Poznan 60-141, Poland., Jeżewska-Frąckowiak J; Department of Molecular Biotechnology, Faculty of Chemistry, University of Gdansk, Gdansk 80-308, Poland.; BioVentures Institute Ltd., Poznan 60-141, Poland., Janus Ł; Department of Molecular Biotechnology, Faculty of Chemistry, University of Gdansk, Gdansk 80-308, Poland.; BioVentures Institute Ltd., Poznan 60-141, Poland., Witkowska M; Department of Molecular Biotechnology, Faculty of Chemistry, University of Gdansk, Gdansk 80-308, Poland., Palczewska M; Department of Molecular Biotechnology, Faculty of Chemistry, University of Gdansk, Gdansk 80-308, Poland., Zylicz-Stachula A; Department of Molecular Biotechnology, Faculty of Chemistry, University of Gdansk, Gdansk 80-308, Poland.; BioVentures Institute Ltd., Poznan 60-141, Poland.
Jazyk:	angličtina
Zdroj:	MethodsX [MethodsX] 2020 Sep 21; Vol. 7, pp. 101070. Date of Electronic Publication: 2020 Sep 21 (Print Publication: 2020).
DOI:	10.1016/j.mex.2020.101070
Abstrakt:	De novo designed bioactive molecules, such as DNA, RNA and peptides, are utilized in increasingly diverse scientific, industrial and biomedical applications. Concatemerization of designed DNA, RNA and peptides may improve their stability, bioactivity and allow for gradual release of the bioactive molecule at the intended destination. In this context, we developed a new method enabling the formation of DNA concatemers for the production of artificial, repetitive genes, encoding concatemeric RNAs and proteins of any nucleotide and amino-acid sequence. The technology recruits the Type IIS SapI restriction endonuclease (REase) for assembling DNA fragments in an ordered head-to-tail-orientation. Alternatively, other commercially available SapI isoschizomers can be used: LguI and thermostable BspQI. Four series of DNA vectors dedicated to the expression of newly formed, concatemeric open reading frames (ORFs), were designed and constructed to meet the technology needs. • Vector-enzymatic DNA fragment amplification technology. • Construction of DNA concatemers many times longer than those available with the use of current de novo gene synthesis methods. • Biosynthesis of protein tandem repeats with programmable function never seen in nature. (© 2020 The Authors. Published by Elsevier B.V.)
Databáze:	MEDLINE
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