{"title":"Assembling DNA Plasmids with the Multi-Kingdom (MK) Cloning System.","authors":"Sarina Scoville, David M Chiasson","doi":"10.1007/978-1-0716-4220-7_26","DOIUrl":null,"url":null,"abstract":"<p><p>The Golden Gate cloning technique is used to assemble DNA parts into higher-order assemblies. Individual parts containing compatible overhangs generated by type IIS restriction enzymes are joined together using DNA ligase. The technique enables users to assemble custom transcription units (TUs) for a wide array of experimental assays. Several Golden Gate cloning systems have been developed; however, they are typically used with a narrow range of organisms. Here we describe the Multi-Kingdom (MK) cloning system that allows users to generate DNA plasmids for use in a broad range of organisms.</p>","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":"2850 ","pages":"467-479"},"PeriodicalIF":0.0000,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Methods in molecular biology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1007/978-1-0716-4220-7_26","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
引用次数: 0
Abstract
The Golden Gate cloning technique is used to assemble DNA parts into higher-order assemblies. Individual parts containing compatible overhangs generated by type IIS restriction enzymes are joined together using DNA ligase. The technique enables users to assemble custom transcription units (TUs) for a wide array of experimental assays. Several Golden Gate cloning systems have been developed; however, they are typically used with a narrow range of organisms. Here we describe the Multi-Kingdom (MK) cloning system that allows users to generate DNA plasmids for use in a broad range of organisms.
金门克隆技术用于将 DNA 部件组装成高阶组件。含有由 IIS 型限制酶产生的兼容悬垂的单个部分通过 DNA 连接酶连接在一起。该技术使用户能够为各种实验检测组装定制的转录单元（TU）。目前已开发出几种金门克隆系统，但它们通常只用于范围较窄的生物。在这里，我们介绍了多王国（MK）克隆系统，它允许用户生成用于多种生物的 DNA 质粒。
期刊介绍：
For over 20 years, biological scientists have come to rely on the research protocols and methodologies in the critically acclaimed Methods in Molecular Biology series. The series was the first to introduce the step-by-step protocols approach that has become the standard in all biomedical protocol publishing. Each protocol is provided in readily-reproducible step-by-step fashion, opening with an introductory overview, a list of the materials and reagents needed to complete the experiment, and followed by a detailed procedure that is supported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice.
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