Abstract：
Inducible gene expression is useful for biotechnological applications and for studying gene regulation and function in bacteria. Many inducible systems that perform in model organisms such as the Gammaproteobacterium Escherichia coli do not perform well in other bacteria that are of biotechnological interest. Typical problems include weak or leaky expression. Here, we describe an invention named ACIT (Alphaproteobacteria chromosomally integrating transcription-control cassette) that is carried on a suicide plasmid to enable insertion into the chromosome of the host. ACIT consists of multiple DNA fragments specifically arranged in a cassette that allows tight transcription control over any gene or gene cluster of interest following homologous recombination. At the heart of the invention is the ability to modify or exchange parts, e.g., promoters, to suit particular bacteria and growth conditions, allowing for customized gene expression control. Furthermore, ACIT provides a basis for a design-build-test approach for controlling gene expression in less studied bacteria. We describe examples of its control over pigment and exopolysaccharide production, growth, cell form, and social behavior in various Alphaproteobacteria.
摘要：
诱导型基因表达可用于生物技术应用和研究细菌中的基因调控和功能。许多可诱导的系统在模型生物如γ变形杆菌大肠杆菌中表现不佳在其他具有生物技术兴趣的细菌中表现不佳。典型的问题包括弱或泄漏的表达。这里,我们描述了一种名为ACIT（染色体整合转录控制盒的α变形杆菌）的发明，该发明被携带在自杀质粒上以能够插入宿主的染色体中。ACIT由多个DNA片段组成,所述DNA片段特异性排列在允许在同源重组后对任何感兴趣的基因或基因簇进行严格转录控制的盒中。本发明的核心是修改或交换零件的能力，例如，promotors,为了适应特定的细菌和生长条件，允许定制的基因表达控制。此外，ACIT为在研究较少的细菌中控制基因表达的设计构建测试方法提供了基础。我们描述了它对色素和胞外多糖生产的控制的例子，增长,细胞形式,和各种阿尔法变形杆菌的社会行为。
