BETA-GLUCURONIDASE (GUS) TRANSPOSONS FOR ECOLOGICAL AND GENETIC-STUDIES OF RHIZOBIA AND OTHER GRAM-NEGATIVE BACTERIA
BETA-GLUCURONIDASE (GUS) TRANSPOSONS FOR ECOLOGICAL AND GENETIC-STUDIES OF RHIZOBIA AND OTHER GRAM-NEGATIVE BACTERIA
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DOI：
10.1099/13500872-141-7-1691
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发表时间：
1995-07-01
期刊：
影响因子：
2.8
通讯作者：
JEFFERSON, RA
中科院分区：
文献类型：
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作者：
WILSON, KJ;SESSITSCH, A;JEFFERSON, RA
描述了一系列转座子，它们含有 gusA 基因，编码 β-葡萄糖醛酸酶 (GUS)，由多种调节型和组成型启动子表达。受调控的启动子包括可被IPTG诱导的tac启动子和在豆科植物根瘤中共生激活的nifH启动子。一个转座子包含具有强 Shine-Dalgarno 翻译起始上下文的 gusA，但没有启动子，因此充当启动子-探针转座子。此外，还描述了大肠杆菌的 gus 操纵子缺失菌株，以及设计用于使用 PFGE 进行染色体作图的转座子。 GUS转座子是在迷你Tn5系统中构建的，可以通过接合转移至革兰氏阴性细菌，并形成稳定的基因组插入。由于植物和许多具有经济重要性的细菌缺乏GUS活性，这些转座子构成了强大的新工具，可用于研究环境中以及与植物相关的细菌的生态学和群体生物学，以及研究这种相互作用的基本分子基础。 GUS 提供多种检测方法，可对标记细菌进行定量检测和空间定位。
A series of transposons are described which contain the gusA gene, encoding beta-glucuronidase (GUS), expressed from a variety of promoters, both regulated and constitutive. The regulated promoters include the tac promoter which can be induced by IPTG, and nifH promoters which are symbiotically activated in legume nodules. One transposon contains gusA with a strong Shine-Dalgarno translation initiation context, but no promoter, and thus acts as a promoter-probe transposon. In addition, a gus operon deletion strain of Escherichia coli, and a transposon designed for use in chromosomal mapping using PFGE, are described. The GUS transposons are constructed in a mini-Tn5 system which can be transferred to Gram-negative bacteria by conjugation, and will form stable genomic insertions. Due to the absence of GUS activity in plants and many bacteria of economic importance, these transposons constitute powerful new tools for studying the ecology and population biology of bacteria in the environment and in association with plants, as well as for studies of the fundamental molecular basis of such interactions. The variety of assays available for GUS enable both quantitative assays and spatial localization of marked bacteria to be carried out.