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Z Pragai

Publications and source records attributed to Z Pragai.

5 recordsLinked to original sources

Essential Bacillus subtilis genes.

To estimate the minimal gene set required to sustain bacterial life in nutritious conditions, we carried out a systematic inactivation of Bacillus subtilis genes. Among approximately 4,100 genes of the organism, only 192 were shown to be indispensable by this or previous work. Another 79 genes were predicted to be essential. The vast majority of essential genes were categorized in relatively few domains of cell metabolism, with about half involved in information processing, one-fifth involved in the synthesis of cell envelope and the determination of cell shape and division, and one-tenth related to cell energetics. Only 4% of essential genes encode unknown functions. Most essential genes are present throughout a wide range of Bacteria, and almost 70% can also be found in Archaea and Eucarya. However, essential genes related to cell envelope, shape, division, and respiration tend to be lost from bacteria with small genomes. Unexpectedly, most genes involved in the Embden-Meyerhof-Parnas pathway are essential. Identification of unknown and unexpected essential genes opens research avenues to better understanding of processes that sustain bacterial life.

Bacillus subtilis↗

Expression of a new operon from Bacillus subtilis, ykzB-ykoL, under the control of the TnrA and PhoP-phoR global regulators.

The ykzB and ykoL genes encode two peptides, of 51 and 60 amino acids, the functions of which are unknown. The ykzB and tnrA genes are contiguous and transcribed divergently. Expression of ykzB and ykoL is induced by glutamate and is under the control of the TnrA global regulator of nitrogen utilization. TnrA regulated its own synthesis in glutamate minimal medium. Two DNA sequences (TnrAB1 and TnrAB2) homologous to the TnrA binding site are present in the region between tnrA and ykzB. Deletion mapping indicated that the TnrAB2 binding site was involved in activation of the ykzB promoter. In addition, transcription of tnrA depends on the presence of the TnrAB1 binding site. The ykzB and ykoL genes are probably in the same transcriptional unit. A single promoter involved in transcription in the presence of glutamate was mapped by primer extension. ykoL expression was induced by phosphate limitation and depended on the PhoP-PhoR two-component regulatory system. Its promoter was mapped to the region between ykoL and ykzB. Four boxes similar to the PhoP binding site are present upstream from the ykoL promoter. These boxes are probably recognized by PhoP approximately P during the activation of transcription in phosphate limitation conditions.

Bacillus subtilis↗

In-vitro selection of porin-deficient mutants of two strains of Klebsiella pneumoniae with reduced susceptibilities to meropenem, but not to imipenem.

We have evaluated the ability of imipenem and meropenem to select, in vitro, resistant mutants of two clinical isolates of Klebsiella pneumoniae producing both SHV and TEM beta-lactamases. Only meropenem selected mutants of both isolates for which the MICs of meropenem, but not imipenem, were markedly higher than those for the parent strains; the MICs of several other beta-lactam antibiotics, including beta-lactam/beta-lactamase inhibitor combinations, for these mutants were also higher than those for the parent strains. In contrast, the MICs for the imipenem-selected mutants were the same as, or similar to, those for the parent strains. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis analysis revealed that an outer membrane protein in both parent strains was absent in the meropenem-selected mutants, but not in the imipenem-selected mutants. This protein is likely to be a porin, the absence of which is presumably associated with impaired beta-lactam permeability and, therefore, the reduced susceptibilities to these antibiotics exhibited by the mutant strains. We believe that this is the first report of the in-vitro selection of porin-deficient mutants of K. pneumoniae following exposure to meropenem.

Drug Resistance, Microbial↗

Investigation of the presence of different broad-spectrum beta-lactamases among clinical isolates of Enterobacteriacae.

Chromosomal or plasmid-encoded beta-lactamases are the most frequent causes of resistance to broad-spectrum beta-lactam antibiotics in clinical isolates of Gram-negative bacteria. Different screening methods can be used for their detection during routine laboratory work, while molecular biological methods may help in the detection of the genetic background of the phenotypic resistance. Clinical isolates of Klebsiella pneumoniae (170) and Enterobacter cloaceae (82) were obtained from different parts of Hungary, whereas those of Serratia marcescens (15) were isolated in our Department from a nosocomial outbreak. Disk diffusion and the Etest were used to screen inducible Class C beta-lactamase and plasmid-mediated extended spectrum beta-lactamases (ESBLs) among clinical isolates of Enterobacteriaceae. Single-strand conformation polymorphism (SSCP) analysis of the PCR products obtained after using SHV-specific primers revealed the presence of SHV-2 and SHV-5 ESBL among 170 K. pneumoniae strains in 12 and 3 cases, respectively. The results of the screening methods and the PCR-SSCP analysis suggested that 14 of the 15 S. marcescens strains not only produced the Class C, inducible chromosomal beta-lactamase, but also acquired a plasmid-mediated SHV-2-type ESBL. One strain isolated from the environment during the outbreak was genetically related to the other isolates, as demonstrated by the different typing methods, but it did not produce ESBL. The in vivo transfer of SHV-2 gene was assumed from an SHV-2 positive K. pneumoniae strain present in the same ward, in the same patient and at the same time. A very high prevalence of the stable derepressed mutants of E. cloaceae was confirmed among the Hungarian isolates. Seventy seven per cent of the strains produced high amounts of beta-lactamase without induction being responsible for their resistance to third-generation cephalosporins. Nineteen per cent of the strains were inducible when cefoxitin or imipenem was used, as confirmed by direct measurement of the MICs with the Etest.

Anti-Bacterial Agents↗