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| ファイル | 記述 | サイズ | フォーマット | |
|---|---|---|---|---|
| s11033-024-09367-y.pdf | 2.33 MB | Adobe PDF | 見る/開く |
| タイトル: | Increase in the solubility of uvsY using a site saturation mutagenesis library for application in a lyophilized reagent for recombinase polymerase amplification |
| 著者: | Morimoto, Kenta Juma, Kevin Maafu Yamagata, Masaya Takita, Teisuke    https://orcid.org/0000-0001-9333-2335 (unconfirmed)Kojima, Kenji Suzuki, Koichiro Yanagihara, Itaru Fujiwara, Shinsuke Yasukawa, Kiyoshi |
| 著者名の別形: | 森本, 健太 山形, 昌也 滝田, 禎亮 保川, 清 |
| キーワード: | Isothermal DNA amplification Recombinase polymerase amplification (RPA) Site saturation mutagenesis library uvsY |
| 発行日: | 27-Feb-2024 |
| 出版者: | Springer Nature |
| 誌名: | Molecular Biology Reports |
| 巻: | 51 |
| 論文番号: | 367 |
| 抄録: | Background: 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. |
| 著作権等: | © The Author(s) 2024 This 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. |
| URI: | http://hdl.handle.net/2433/290350 |
| DOI(出版社版): | 10.1007/s11033-024-09367-y |
| PubMed ID: | 38411701 |
| 出現コレクション: | 学術雑誌掲載論文等 |
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