Rajendran N, Jahn D, Jayaraman K, Marahiel M A
Biochemie/Fachbereich Chemie, Philipps-Universität Marburg, FRG.
FEMS Microbiol Lett. 1994 Jan 15;115(2-3):191-6. doi: 10.1111/j.1574-6968.1994.tb06636.x.
Pseudomonas fluorescens was subjected to insertion mutagenesis studies using the transposon Tn5-GM to generate mutants deficient in antibacterial activity minus mutants. The transposon located on the temperature-sensitive plasmid pCHR84 was conjugally transferred into the non-pathogenic pseudomonad using the triparental mating procedure. Random integration of Tn5-GM into the chromosome of P. fluorescens was achieved by heat treatment of the transformed cells at 42 degrees C. Approximately 2% of transconjugants revealed an auxotrophic phenotype indicating efficient integration of the employed transposon into the chromosome of P. fluorescens. One transposon insertion mutant was obtained showing an antibacterial activity minus phenotype. This mutant (MM-7) was found to be defective in the production of an unidentified antibacterial compound against B. subtilis. These results introduce Tn5 transposon mutagenesis as a new useful tool for the molecular analysis of P. fluorescens.
使用转座子Tn5-GM对荧光假单胞菌进行插入诱变研究，以产生抗菌活性缺陷型突变体（负突变体）。位于温度敏感质粒pCHR84上的转座子通过三亲本杂交程序接合转移到非致病性假单胞菌中。通过在42℃对转化细胞进行热处理，实现Tn5-GM随机整合到荧光假单胞菌的染色体中。约2%的转接合子表现出营养缺陷型表型，表明所用转座子有效整合到荧光假单胞菌的染色体中。获得了一个表现出抗菌活性缺陷型表型的转座子插入突变体。发现该突变体（MM-7）在产生针对枯草芽孢杆菌的一种未鉴定抗菌化合物方面存在缺陷。这些结果表明Tn5转座子诱变是荧光假单胞菌分子分析的一种新的有用工具。
