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モトハシ タケシ
MOTOHASHI TAKESHI
本橋 健 所属 京都産業大学 生命科学部 先端生命科学科 職種 教授 |
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| 言語種別 | 英語 |
| 発行・発表の年月 | 2017 |
| 形態種別 | 研究論文 |
| 査読 | 査読あり |
| 標題 | Seamless ligation cloning extract (SLiCE) method using cell lysates from laboratory escherichia coli strains and its application to slip site-directed mutagenesis |
| 執筆形態 | その他 |
| 掲載誌名 | Methods in Molecular Biology |
| 出版社・発行元 | Methods in Molecular Biology |
| 巻・号・頁 | 1498,pp.349-357 |
| 著者・共著者 | Motohashi, K. |
| 概要 | Cell lysates from laboratory Escherichia coli strains endogenously exhibit homologous recombination activity, which can be utilized for seamless DNA cloning in vitro. This method, termed S eamless L igation C loning E xtract (SLiCE) cloning, enables high cloning efficiency with simultaneous integration of two unpurified DNA fragments into a vector. In addition, the SLiCE method is highly cost-effective, as several laboratory E. coli strains may be utilized as sources of SLiCE. Previously, the SLiCE technique has been applied to sitedirected mutagenesis to develop a novel technique termed SLiCE-mediated polymerase chain reaction (PCR)-based site-directed mutagenesis (SLiP site-directed mutagenesis). Two DNA fragments containing a mutation site can be simultaneously integrated into a vector while avoiding the introduction of undesirable mutations in the vector. Therefore, SLiP site-directed mutagenesis simplifies multiple procedures involved in PCR-based site-directed mutagenesis such as overlap extension method PCR or the Megaprimer method. |
| DOI | 10.1007/978-1-4939-6472-7_23 |
| ISSN | 1064-3745 |
| PMID | 27709587 |
| Put Code(ORCID) | 37642199 |
| PermalinkURL | http://www.scopus.com/inward/record.url?eid=2-s2.0-84990924169&partnerID=MN8TOARS |