恶臭假单胞菌中基因融合的体内形成以及用于直接亚克隆Mu d1和Mu d2融合体的通用广宿主载体的构建。
In vivo formation of gene fusions in Pseudomonas putida and construction of versatile broad-host-range vectors for direct subcloning of Mu d1 and Mu d2 fusions.
作者信息
Simon V, Schumann W
机构信息
Lehrstuhl für Genetik, Universität Bayreuth, Federal Republic of Germany.
出版信息
Appl Environ Microbiol. 1987 Jul;53(7):1649-54. doi: 10.1128/aem.53.7.1649-1654.1987.
Abstract
The Mu d1 and Mu d2 prophages were integrated into the conjugative broad-host-range plasmid R751. The two plasmids were then transferred into Pseudomonas putida, and derivatives carrying intact Mu prophages were recovered. After induction of Mu at 42 degrees C, both operon and gene fusions were observed on 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-Gal) plates. Broad-host-range vectors were constructed which allow direct cloning of both operon or gene fusions and their analysis in Escherichia coli and P. putida. By using one of these vectors, two operon fusions were isolated from the P. putida chromosome and comparatively analyzed in E. coli and P. putida.
摘要
Mu d1和Mu d2原噬菌体被整合到接合型广宿主范围质粒R751中。然后将这两种质粒转移到恶臭假单胞菌中，并回收携带完整Mu原噬菌体的衍生物。在42℃诱导Mu后，在5-溴-4-氯-3-吲哚基-β-D-吡喃半乳糖苷（X-Gal）平板上观察到操纵子融合和基因融合。构建了广宿主范围载体，可直接克隆操纵子或基因融合体，并在大肠杆菌和恶臭假单胞菌中进行分析。通过使用其中一种载体，从恶臭假单胞菌染色体中分离出两个操纵子融合体，并在大肠杆菌和恶臭假单胞菌中进行比较分析。
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