Gruber Steffen, Schwab Helmut, Koefinger Petra
Institute of Molecular Biotechnology, Graz University of Technology, NAWI Graz, Petersgasse 14, 8010 Graz, Austria.
Institute of Molecular Biotechnology, Graz University of Technology, NAWI Graz, Petersgasse 14, 8010 Graz, Austria.
N Biotechnol. 2015 Dec 25;32(6):552-8. doi: 10.1016/j.nbt.2015.03.015. Epub 2015 Apr 9.
The Gram-negative bacterium Escherichia coli is currently the most efficient and widely used prokaryotic host for recombinant protein and metabolite production. However, due to some limitations and to various interesting features of other Gram-negative bacteria efficient vector systems applicable to a broad range are desired. Basic building blocks for plasmid-based vectors include besides the need for a suitable selection marker in the first line a proper replication and maintenance system. In addition to these basic requirements, further elements are needed for Gram-negative bacteria beyond E. coli, such as Pseudomonas pudita, Ralstonia eutropha, Burkholderia glumae or Acinetobacter sp.. Established building blocks have to be adapted and new building blocks providing the desired functions need to be identified and exploited. This minireview addresses so far described and used genetic elements for broad host range replication, efficient plasmid maintenance, and conjugative plasmid transfer as well as expression elements and protein secretion signals. The industrially important bacterium R. eutropha H16 was chosen as a model organism to provide specific data on the effectivity and utility of building blocks based on such genetic elements.
革兰氏阴性菌大肠杆菌是目前用于重组蛋白和代谢产物生产的最有效且应用最广泛的原核宿主。然而，由于存在一些局限性以及其他革兰氏阴性菌具有各种有趣的特性，人们期望获得适用于广泛范围的高效载体系统。基于质粒的载体的基本组成部分，首先除了需要合适的选择标记外，还需要适当的复制和维持系统。除了这些基本要求外，对于大肠杆菌以外的革兰氏阴性菌，如恶臭假单胞菌、嗜麦芽窄食单胞菌、水稻细菌性谷枯病菌或不动杆菌属，还需要其他元件。已有的组成部分必须进行调整，并且需要识别和利用能提供所需功能的新组成部分。本综述讨论了迄今为止已描述和使用的用于广泛宿主范围复制、高效质粒维持、接合质粒转移的遗传元件，以及表达元件和蛋白质分泌信号。选择具有工业重要性的嗜麦芽窄食单胞菌H16作为模式生物，以提供基于此类遗传元件的组成部分的有效性和实用性的具体数据。
