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pGinger 表达质粒家族。

The pGinger Family of Expression Plasmids.

机构信息

Joint BioEnergy Institute, Emeryville, California, USA.

Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, California, USA.

出版信息

Microbiol Spectr. 2023 Jun 15;11(3):e0037323. doi: 10.1128/spectrum.00373-23. Epub 2023 May 22.

Abstract

The pGinger suite of expression plasmids comprises 43 plasmids that will enable precise constitutive and inducible gene expression in a wide range of Gram-negative bacterial species. Constitutive vectors are composed of 16 synthetic constitutive promoters upstream of red fluorescent protein (RFP), with a broad-host-range BBR1 origin and a kanamycin resistance marker. The family also has seven inducible systems (Jungle Express, Psal/NahR, Pm/XylS, Prha/RhaS, LacO1/LacI, LacUV5/LacI, and Ptet/TetR) controlling RFP expression on BBR1/kanamycin plasmid backbones. For four of these inducible systems (Jungle Express, Psal/NahR, LacO1/LacI, and Ptet/TetR), we created variants that utilize the RK2 origin and spectinomycin or gentamicin selection. Relevant RFP expression and growth data have been collected in the model bacterium Escherichia coli as well as Pseudomonas putida. All pGinger vectors are available via the Joint BioEnergy Institute (JBEI) Public Registry. Metabolic engineering and synthetic biology are predicated on the precise control of gene expression. As synthetic biology expands beyond model organisms, more tools will be required that function robustly in a wide range of bacterial hosts. The pGinger family of plasmids constitutes 43 plasmids that will enable both constitutive and inducible gene expression in a wide range of nonmodel .

摘要

pGinger 表达质粒套件包含 43 个质粒,可在多种革兰氏阴性细菌物种中实现精确的组成型和诱导型基因表达。组成型载体由红色荧光蛋白 (RFP) 上游的 16 个合成组成型启动子组成,具有广泛宿主范围的 BBR1 原点和卡那霉素抗性标记。该家族还具有七个诱导系统(Jungle Express、Psal/NahR、Pm/XylS、Prha/RhaS、LacO1/LacI、LacUV5/LacI 和 Ptet/TetR),控制 BBR1/卡那霉素质粒骨架上的 RFP 表达。对于其中四个诱导系统(Jungle Express、Psal/NahR、LacO1/LacI 和 Ptet/TetR),我们创建了利用 RK2 原点和壮观霉素或庆大霉素选择的变体。在模型细菌大肠杆菌和恶臭假单胞菌中已经收集了相关的 RFP 表达和生长数据。所有 pGinger 载体均可通过联合生物能源研究所 (JBEI) 公共注册处获得。代谢工程和合成生物学基于基因表达的精确控制。随着合成生物学超越模式生物,将需要更多在广泛的细菌宿主中稳健运行的工具。pGinger 质粒家族由 43 个质粒组成,可在多种非模型细菌中实现组成型和诱导型基因表达。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c256/10269703/7e56e16a29f5/spectrum.00373-23-f001.jpg

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