Department of Biology, Villanova University, Villanova, PA 19085, USA.
J Microbiol Biotechnol. 2010 May;20(5):871-4.
Many studies require expression analysis of the same gene/promoter across a range of bacterial genera. However, there is currently a lack of availability of reporters based on the broad host range IncQ replicon, which is compatible with a popular improved IncP transfer system that is self-transfer defective. We report IncQ lacZ reporter plasmids with features including: (1) compatibility with IncP, IncW, and pBHR/pBBR replicons, (2) a variety of antibiotic markers (Sp-r, Sm-r, Km-r, Cm-r), (3) convenient mobilization via a novel self-transfer-defective IncP conjugation system, and (4) complete DNA sequences. Utility is demonstrated using three different promoters in different Gram negative genera.
许多研究都需要对广泛的细菌属中的同一基因/启动子进行表达分析。然而,目前缺乏基于广谱宿主范围 IncQ 复制子的报告基因,而 IncQ 复制子与一种流行的改良 IncP 转移系统兼容,该系统本身的转移是有缺陷的。我们报告了带有以下特征的 IncQ lacZ 报告质粒:(1) 与 IncP、IncW 和 pBHR/pBBR 复制子兼容,(2) 多种抗生素标记(Sp-r、Sm-r、Km-r、Cm-r),(3) 通过新型的自我转移缺陷 IncP 共轭系统方便地进行动员,以及 (4) 完整的 DNA 序列。通过革兰氏阴性菌中不同属的三个不同启动子证明了其实用性。