Tanaka K, Takahashi H
Institute of Molecular and Cellular Biosciences, University of Tokyo, Japan.
Gene. 1994 Dec 2;150(1):81-5. doi: 10.1016/0378-1119(94)90862-1.
A homologue of the rpoS gene of Escherichia coli was cloned from Pseudomonas aeruginosa PAO1 by hybridization with an oligodeoxyribonucleotide probe designed from an amino-acid stretch conserved among the principal sigma factors of eubacteria. Two open reading frames, the pcm gene and the orf-297 of unknown function, were found in the upstream region of rpoS, and in the same order as in E. coli. The rpoS gene of P. aeruginosa was expressed in E. coli and complemented the catalase deficiency of the rpoS mutant of E. coli. The RpoS protein of P. aeruginosa was identified by Western blot analysis in both P. aeruginosa (Pa) and the transformed E. coli. Levels of RpoS of Pa increased drastically at the onset of the stationary growth phase.
通过与基于真细菌主要σ因子中保守氨基酸序列设计的寡脱氧核糖核苷酸探针杂交，从铜绿假单胞菌PAO1中克隆出大肠杆菌rpoS基因的同源物。在rpoS的上游区域发现了两个开放阅读框，即功能未知的pcm基因和orf - 297，其排列顺序与大肠杆菌相同。铜绿假单胞菌的rpoS基因在大肠杆菌中表达，并弥补了大肠杆菌rpoS突变体的过氧化氢酶缺陷。通过蛋白质免疫印迹分析在铜绿假单胞菌（Pa）和转化的大肠杆菌中均鉴定出了铜绿假单胞菌的RpoS蛋白。在稳定生长期开始时，Pa的RpoS水平急剧增加。
