Climent N, Ferrer S, Rubires X, Merino S, Tomás J M, Regué M
Department of Microbiology and Parasitology, Faculty of Pharmacy, University of Barcelona, Spain.
Res Microbiol. 1997 Feb;148(2):133-43. doi: 10.1016/S0923-2508(97)87644-9.
A cosmid-based genomic library of Klebsiella pneumoniae 52145 (O1:K2) was introduced into Escherichia coli, and clones were screened for the bacteriocin 28b resistance phenotype. One clone was found which conferred partial resistance to bacteriocin 28b. By using Tn5tac1 insertions, it was shown that this phenotype was due to the expression, in E. coli, of an outer-membrane protein (OMP) with an apparent molecular mass of 17 kDa (OmpK17). The DNA region defined by insertion mutagenesis was sequenced and found to contain an ORF of 510 bp. The deduced amino acid sequence has 170 residues with a theoretical molecular mass of 18.4 kDa. The protein contains an N-terminal signal sequence of 24 amino acid residues. When compared with other enterobacterial OMPs, OmpK17 most closely resembles members of a family of small OMPs of Enterobacteriaceae the known functions of which appear to be related to virulence. Immunoblotting experiments showed that OmpK17 is also present in various K. pneumoniae strains belonging to different O and K serotypes.
将肺炎克雷伯菌52145(O1:K2)基于黏粒的基因组文库导入大肠杆菌,并筛选对细菌素28b具有抗性表型的克隆。发现一个克隆赋予了对细菌素28b的部分抗性。通过使用Tn5tac1插入,表明该表型是由于在大肠杆菌中表达了一种表观分子量为17 kDa的外膜蛋白(OmpK17)。对由插入诱变定义的DNA区域进行测序,发现其包含一个510 bp的开放阅读框。推导的氨基酸序列有170个残基,理论分子量为18.4 kDa。该蛋白含有一个24个氨基酸残基的N端信号序列。与其他肠杆菌外膜蛋白相比,OmpK17与肠杆菌科小外膜蛋白家族的成员最为相似,已知其功能似乎与毒力有关。免疫印迹实验表明,OmpK17也存在于属于不同O和K血清型的各种肺炎克雷伯菌菌株中。