Lawes M, Maloy S
Department of Microbiology, University of Illinois, Urbana 61801.
J Bacteriol. 1995 Mar;177(5):1383-7. doi: 10.1128/jb.177.5.1383-1387.1995.
The transposable bacteriophage Mu and its mini-Mu derivatives are useful tools for the genetic analysis of many bacteria. A variety of antibiotic-resistant Mu derivatives have been constructed, allowing direct selection for cells which contain the transposon. However, in many cases a counterselection against the transposon would greatly facilitate further genetic analysis. In this paper we report the construction of MudSacI, a mini-Mu derived transposon containing the sacB (secretory levansucrase) gene of Bacillus subtilis, which confers sucrose sensitivity upon gram-negative bacteria. We describe the use of this transposon as a tool for rapid genetic mapping of chromosomal genes in Salmonella typhimurium. Simple modifications of this approach should facilitate rapid mapping in many other bacteria as well.
可转座噬菌体Mu及其微型Mu衍生物是对许多细菌进行遗传分析的有用工具。已经构建了多种抗抗生素的Mu衍生物，从而能够直接筛选含有转座子的细胞。然而，在许多情况下，针对转座子的反选择将极大地促进进一步的遗传分析。在本文中，我们报告了MudSacI的构建，它是一种源自微型Mu的转座子，含有枯草芽孢杆菌的sacB（分泌型果聚糖蔗糖酶）基因，该基因赋予革兰氏阴性菌蔗糖敏感性。我们描述了将这种转座子用作鼠伤寒沙门氏菌染色体基因快速遗传定位工具的用途。对该方法进行简单修改也应有助于在许多其他细菌中进行快速定位。
