Protecting Linear DNA Templates in Cell-Free Expression Systems from Diverse Bacteria

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dc.contributor.authorYim, Sung Sunko
dc.contributor.authorJohns, Nathan, Iko
dc.contributor.authorNoireaux, Vincentko
dc.contributor.authorWang, Harris H.ko
dc.date.accessioned2022-07-20T02:00:23Z-
dc.date.available2022-07-20T02:00:23Z-
dc.date.created2022-07-20-
dc.date.issued2020-10-
dc.identifier.citationACS SYNTHETIC BIOLOGY, v.9, no.10, pp.2851 - 2855-
dc.identifier.issn2161-5063-
dc.identifier.urihttp://hdl.handle.net/10203/297407-
dc.description.abstractRecent advances in cell-free systems have opened up new capabilities in synthetic biology from rapid prototyping of genetic circuits and metabolic pathways to portable diagnostics and biomanufacturing. A current bottleneck in cell-free systems, especially those employing non-E. coli bacterial species, is the required use of plasmid DNA, which can be laborious to construct, clone, and verify. Linear DNA templates offer a faster and more direct route for many cell-free applications, but they are often rapidly degraded in cell-free reactions. In this study, we evaluated GamS from lambda-phage, DNA fragments containing Chi-sites, and Ku from Mycobacterium tuberculosis for their ability to protect linear DNA templates in diverse bacterial cell-free systems. We show that these nuclease inhibitors exhibit differential protective activities against endogenous exonucleases in five different cell-free lysates, highlighting their utility for diverse bacterial species. We expect these linear DNA protection strategies will accelerate high-throughput approaches in cell-free synthetic biology.-
dc.languageEnglish-
dc.publisherAMER CHEMICAL SOC-
dc.titleProtecting Linear DNA Templates in Cell-Free Expression Systems from Diverse Bacteria-
dc.typeArticle-
dc.identifier.wosid000582582100023-
dc.identifier.scopusid2-s2.0-85096590035-
dc.type.rimsART-
dc.citation.volume9-
dc.citation.issue10-
dc.citation.beginningpage2851-
dc.citation.endingpage2855-
dc.citation.publicationnameACS SYNTHETIC BIOLOGY-
dc.identifier.doi10.1021/acssynbio.0c00277-
dc.contributor.localauthorYim, Sung Sun-
dc.contributor.nonIdAuthorJohns, Nathan, I-
dc.contributor.nonIdAuthorNoireaux, Vincent-
dc.contributor.nonIdAuthorWang, Harris H.-
dc.description.isOpenAccessN-
dc.type.journalArticleArticle-
dc.subject.keywordAuthorcell-free expression systems-
dc.subject.keywordAuthorlinear DNA-
dc.subject.keywordAuthorRecBCD-
dc.subject.keywordAuthorGamS-
dc.subject.keywordAuthorChi-
dc.subject.keywordAuthorKu-
dc.subject.keywordPlusREPAIR-
dc.subject.keywordPlusPROTEIN-
dc.subject.keywordPlusSTABILITY-
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