Cargando…
HyperXpress: Rapid Single Vessel DNA Assembly and Protein Production in Microliterscale
Rapid prototyping of biological functions has the common aim of generating, screening, and selecting variant libraries as quickly as possible. This approach is now to be extended by the HyperXpress workflow, which connects ligase cycling reaction for DNA assembly, multiply-primed rolling circle ampl...
Autores principales: | , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9011061/ https://www.ncbi.nlm.nih.gov/pubmed/35433646 http://dx.doi.org/10.3389/fbioe.2022.832176 |
_version_ | 1784687611475918848 |
---|---|
author | Zibulski, Darius Leon Schlichting, Niels Kabisch, Johannes |
author_facet | Zibulski, Darius Leon Schlichting, Niels Kabisch, Johannes |
author_sort | Zibulski, Darius Leon |
collection | PubMed |
description | Rapid prototyping of biological functions has the common aim of generating, screening, and selecting variant libraries as quickly as possible. This approach is now to be extended by the HyperXpress workflow, which connects ligase cycling reaction for DNA assembly, multiply-primed rolling circle amplification for signal amplification, and cell-free protein synthesis to a single vessel reaction in the lower µl scale. After substantial optimization of the method a proof-of-principle demonstrating the high flexibility of HyperXpress for semi-rational protein engineering by expanding, reducing, and replacing β-strands of three different green fluorescent proteins is described. These single-day experiments resulted in six functional, new-to-nature GFP prototypes. |
format | Online Article Text |
id | pubmed-9011061 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-90110612022-04-16 HyperXpress: Rapid Single Vessel DNA Assembly and Protein Production in Microliterscale Zibulski, Darius Leon Schlichting, Niels Kabisch, Johannes Front Bioeng Biotechnol Bioengineering and Biotechnology Rapid prototyping of biological functions has the common aim of generating, screening, and selecting variant libraries as quickly as possible. This approach is now to be extended by the HyperXpress workflow, which connects ligase cycling reaction for DNA assembly, multiply-primed rolling circle amplification for signal amplification, and cell-free protein synthesis to a single vessel reaction in the lower µl scale. After substantial optimization of the method a proof-of-principle demonstrating the high flexibility of HyperXpress for semi-rational protein engineering by expanding, reducing, and replacing β-strands of three different green fluorescent proteins is described. These single-day experiments resulted in six functional, new-to-nature GFP prototypes. Frontiers Media S.A. 2022-04-01 /pmc/articles/PMC9011061/ /pubmed/35433646 http://dx.doi.org/10.3389/fbioe.2022.832176 Text en Copyright © 2022 Zibulski, Schlichting and Kabisch. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Bioengineering and Biotechnology Zibulski, Darius Leon Schlichting, Niels Kabisch, Johannes HyperXpress: Rapid Single Vessel DNA Assembly and Protein Production in Microliterscale |
title | HyperXpress: Rapid Single Vessel DNA Assembly and Protein Production in Microliterscale |
title_full | HyperXpress: Rapid Single Vessel DNA Assembly and Protein Production in Microliterscale |
title_fullStr | HyperXpress: Rapid Single Vessel DNA Assembly and Protein Production in Microliterscale |
title_full_unstemmed | HyperXpress: Rapid Single Vessel DNA Assembly and Protein Production in Microliterscale |
title_short | HyperXpress: Rapid Single Vessel DNA Assembly and Protein Production in Microliterscale |
title_sort | hyperxpress: rapid single vessel dna assembly and protein production in microliterscale |
topic | Bioengineering and Biotechnology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9011061/ https://www.ncbi.nlm.nih.gov/pubmed/35433646 http://dx.doi.org/10.3389/fbioe.2022.832176 |
work_keys_str_mv | AT zibulskidariusleon hyperxpressrapidsinglevesseldnaassemblyandproteinproductioninmicroliterscale AT schlichtingniels hyperxpressrapidsinglevesseldnaassemblyandproteinproductioninmicroliterscale AT kabischjohannes hyperxpressrapidsinglevesseldnaassemblyandproteinproductioninmicroliterscale |