Stapleton M J, Jagger K S, Warren R L
J Bacteriol. 1984 Jan;157(1):7-12. doi: 10.1128/jb.157.1.7-12.1984.
Transposon Tn5 was used to generate protease-deficient insertion mutants of Pseudomonas aeruginosa. The presence of Tn5 in the chromosome of P. aeruginosa was demonstrated by transduction and DNA-DNA hybridization. The altered protease production and kanamycin resistance were cotransduced into a wild-type P. aeruginosa strain. A radiolabeled probe of Tn5 DNA hybridized to specific BamHI fragments isolated from the insertion mutants. Two independently isolated Tn5 insertion mutants had reduced protease production, partially impaired elastase activity, and no immunologically reactive alkaline protease.
转座子Tn5被用于构建铜绿假单胞菌蛋白酶缺陷型插入突变体。通过转导和DNA-DNA杂交证明了Tn5存在于铜绿假单胞菌的染色体中。蛋白酶产生的改变和卡那霉素抗性被共转导到野生型铜绿假单胞菌菌株中。Tn5 DNA的放射性标记探针与从插入突变体中分离出的特定BamHI片段杂交。两个独立分离的Tn5插入突变体的蛋白酶产生减少,弹性蛋白酶活性部分受损,且没有免疫反应性碱性蛋白酶。