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dc.contributor.authorMorimoto, Kentaen
dc.contributor.authorJuma, Kevin Maafuen
dc.contributor.authorYamagata, Masayaen
dc.contributor.authorTakita, Teisukeen
dc.contributor.authorKojima, Kenjien
dc.contributor.authorSuzuki, Koichiroen
dc.contributor.authorYanagihara, Itaruen
dc.contributor.authorFujiwara, Shinsukeen
dc.contributor.authorYasukawa, Kiyoshien
dc.contributor.alternative森本, 健太ja
dc.contributor.alternative山形, 昌也ja
dc.contributor.alternative滝田, 禎亮ja
dc.contributor.alternative保川, 清ja
dc.date.accessioned2024-11-15T05:32:53Z-
dc.date.available2024-11-15T05:32:53Z-
dc.date.issued2024-02-27-
dc.identifier.urihttp://hdl.handle.net/2433/290350-
dc.description.abstractBackground: Recombinase uvsY from bacteriophage T4, along with uvsX, is a key enzyme for recombinase polymerase amplification (RPA), which is used to amplify a target DNA sequence at a constant temperature. uvsY, though essential, poses solubility challenges, complicating the lyophilization of RPA reagents. This study aimed to enhance uvsY solubility. Methods: Our hypothesis centered on the C-terminal region of uvsY influencing solubility. To test this, we generated a site-saturation mutagenesis library for amino acid residues Lys91-Glu134 of the N-terminal (His)6-tagged uvsY. Results: Screening 480 clones identified A116H as the variant with superior solubility. Lyophilized RPA reagents featuring the uvsY variant A116H demonstrated enhanced performance compared to those with wild-type uvsY. Conclusions: The uvsY variant A116H emerges as an appealing choice for RPA applications, offering improved solubility and heightened lyophilization feasibility.en
dc.language.isoeng-
dc.publisherSpringer Natureen
dc.rights© The Author(s) 2024en
dc.rightsThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.en
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/-
dc.subjectIsothermal DNA amplificationen
dc.subjectRecombinase polymerase amplification (RPA)en
dc.subjectSite saturation mutagenesis libraryen
dc.subjectuvsYen
dc.titleIncrease in the solubility of uvsY using a site saturation mutagenesis library for application in a lyophilized reagent for recombinase polymerase amplificationen
dc.typejournal article-
dc.type.niitypeJournal Article-
dc.identifier.jtitleMolecular Biology Reportsen
dc.identifier.volume51-
dc.relation.doi10.1007/s11033-024-09367-y-
dc.textversionpublisher-
dc.identifier.artnum367-
dc.identifier.pmid38411701-
dcterms.accessRightsopen access-
dc.identifier.pissn0301-4851-
dc.identifier.eissn1573-4978-
出現コレクション:学術雑誌掲載論文等

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