通用联合模块质粒：在任何微生物宿主中进行 Golden Gate 组装的灵活平台。

Joint Universal Modular Plasmids: A Flexible Platform for Golden Gate Assembly in Any Microbial Host.

机构信息

School of Biological Sciences, University of Edinburgh, Edinburgh, UK.

Zhejiang University-University of Edinburgh Joint Research Centre for Engineering Biology, Zhejiang University, Haining, China.

出版信息

Methods Mol Biol. 2020;2205:255-273. doi: 10.1007/978-1-0716-0908-8_15.

DOI:10.1007/978-1-0716-0908-8_15

PMID:32809204

Abstract

Modular cloning standards based on Golden Gate DNA assembly allow for construction of complex DNA constructs over several rounds of assembly. Despite being reliable and automation-friendly, each standard uses a specific set of vectors, requiring researchers to generate new tool kits for novel hosts and cloning applications. JUMP vectors (Valenzuela-Ortega and French, bioRxiv 799585, 2019) combine the robustness of modular cloning standards with the Standard European Vector Architecture and a flexible design that allows researchers to easily modify the vector backbone via secondary cloning sites. This flexibility allows for JUMP vectors to be used in a wide variety of applications and hosts.

摘要

基于 Golden Gate DNA 组装的模块化克隆标准允许在多个组装循环中构建复杂的 DNA 构建体。尽管这种方法可靠且易于自动化，但每个标准都使用特定的载体集，这要求研究人员为新的宿主和克隆应用生成新的工具包。JUMP 载体（Valenzuela-Ortega 和 French，bioRxiv 799585，2019）将模块化克隆标准的稳健性与标准欧洲载体架构以及灵活的设计相结合，允许研究人员通过二级克隆位点轻松修改载体骨架。这种灵活性使得 JUMP 载体能够在各种应用和宿主中使用。