Effect of R-plasmid RP1 and nutrient depletion on the gross cellular composition of Escherichia coli and its resistance to some uncoupling phenols.

The resistance of Escherichia coli batch cultures depleted of carbon (C-dep), magnesium (Mg-dep), or phosphate (P-dep) against low concentrations of 3-chlorophenol, 4-chlorophenol, or 2-phenoxyethanol varied. C-dep cultures were always significantly more sensitive than Mg-dep or P-dep cultures. The presence of R-plasmid RP1 increased the sensitivity of C-dep cultures to 3- and 4-chlorophenol, yet had little effect on those cultured depleted in magnesium or phosphate ions. Cultures with R-plasmid RP1 had increased levels of beta-polyhydroxybutyrate irrespective of the nature of the depleting nutrient. P-dep bacteria had less than one-third of the phospholipid of other cell types, this deficiency being compensated for by increases in fatty acid and neutral lipid content. The reduction in phospholipid content of P-dep cultures was entirely accounted for by decreased diphosphatidylglycerol and phosphatidylethanolamine levels in these cells.     

Effect of R-plasmid RP1 on surface hydrophobicity of Proteus mirabilis

The presence of R-plasmid RP1, as well as the conditions of growth, affected the surface hydrophobicity of a clinical isolate of Proteus mirabilis. However, results depended upon the method of assessment. Stationary phase plasmid-containing cells appeared to be less hydrophobic than plasmid-free cells when hydrophobicity was measured by the contact angle method, but more hydrophobic when measured by bacterial adherence to hydrocarbons or hydrophobic interaction chromatography. Cells growing in a chemostat differed in hydrophobicity from stationary phase cells and results varied with the growth rate. Plasmid-mediated effects were greatest in iron-depleted cells, and differences between plasmid-containing and plasmid-free cells were virtually eliminated by pre-treatment with antiserum.     

Resistance of Escherichia coli to tetracyclines

1. A strain of Escherichia coli highly resistant to chlortetracycline and partially cross-resistant to tetracycline has been isolated. 2. The nitro-reductase system of the resistant cells was inhibited to a smaller extent by chlortetracycline than was the corresponding enzyme of sensitive cells. 3. The incorporation of leucine in vitro into the ribosomal protein of cell-free preparations from sensitive and resistant cells was equally inhibited by chlortetracycline. 4. Resistant cells accumulated much less chlortetracycline and tetracycline than did sensitive cells when both were cultured in the presence of these drugs. 5. The uptake of tetracycline by both sensitive and resistant E. coli was dependent on the presence of glucose in the medium. 6. Fractionation of cells cultured in medium containing [¹⁴C]chlortetracycline indicated that the largest proportion of radioactivity in sensitive cells was in the fraction consisting mainly of cell-wall material. There was no concentration of radioactivity in any one fraction of the resistant cells. 7. No evidence could be obtained for a specific tetracycline-excretion system in the resistant cells. 8. The significance of these results in relation to current theories of the antibiotic action of and resistance to the tetracycline drugs is discussed.     

The Influence of Temperature on the Effect of Bacteriostatic Concentrations of Phenol Against Escherichia coli

S ummary . During early exponential growth of Escherichia coli in the absence of phenol there is a natural death rate at 20, 30, and 44° but at the optimum temperature around 37° there is little if any significant death. The influence of a rise in temperature from 20 to 44° is to decrease the generation time and at 44° the lower generation time compensates for a reduced generation index. The main effect of sub-bacteriostatic concentrations of phenol is to increase the generation time but at 30, 37 and 44° there is a significant reduction in the generation index at the higher concentrations resulting in a dynamic bacteriostasis. At 20° bacteriostasis is due mainly to a large generation time but there is a little growth and so bacteriostasis is essentially dynamic. There is also evidence to suggest that the effect of a particular concentration of phenol on the generation index is not merely influenced by the temperature but by the generation time under the particular set of conditions. If phenol is added to rapidly growing cultures of E. coli the effect of a rise in temperature is to reduce the concentration required for bacteriostasis but if it is added during the lag phase there is a maximum in the bacteriostatic concentration between 20 and 37°.     

Escherichia coli strains isolated from surface waters. Distribution by resistance to antibiotics and R-plasmid transfer

Samples were taken from surface waters in Csongrád county in the months March through December, 1980. Escherichia coli isolates selected at random were tested for resistance to 5 antibiotics and for R-plasmid carriership. Of the strains isolated from Tisza river at each of 8 sampling sites, 50-60% were sensitive to all the 5 antibiotics. The percentage of sensitivity was much lower if only the strains isolated during the summer months, when the water level was high, or those isolated from affluents and backwaters were taken into account. The frequency of resistance was the highest for tetracycline, followed in order by ampicillin, streptomycin, kanamycin and chloramphenicol. R-plasmid was carried by 43% of the resistant isolates tested, mainly by multiresistant ones.     

Expression of a novel R-plasmid pEB017 compared to pKM101 in Escherichia coli wild-type, recA and uvrA strains

The expression of bacterial resistance to UV irradiation and nitrofurantoin by a novel R-plasmid pEB017 in DNA-repair-proficient (wild-type) and -deficient (recA; uvrA) host strains was compared to the effects of plasmid pKM101 in the isogenic strains. pEB017 partially protected the uvrA strain, and completely protected the wild-type and recA strains from the killing effect of UV irradiation; pKM101 had no effect on the recA strain and only enhanced the survival of the wild-type and the uvrA strains after UV irradiation. pEB017 conferred nitrofurantoin resistance 10-fold on the wild-type and the recA strains and 4-fold on the uvrA strain; pKM101 did not confer nitrofurantoin resistance on the wild-type and recA strains but gave 4-fold resistance in the uvrA strain.     

Resistance to the peptide-like antibiotic negamycin in Escherichia coli

An $\text{Escherichia coli}$ mutant resistant to the peptide-like antibiotic negamycin carries a mutation, NEG40, which maps at minute 65 on the bacterial genome. Termination of protein synthesis, which is normally blocked by negamycin in wild type cellular extracts, continues with cellular extracts from the mutant in the presence of the drug; indeed, release of complete peptides from the polysomes still proceeds over a wide range of drug concentrations. The data suggest that the NEG40 mutation affects one of the components of the termination complex (ribosome or release factor).     

Resistance Transfer to the Resident Intestinal Escherichia coli of Rats

The transfer of resistance factors from introduced donor cells to the resident intestinal Escherichia coli flora of conventional rats was tested.     

Host dependence of RP1-specified resistance to ampicillin: differential expression in Escherichia coli and Rhizobium leguminosarum.

Rhizobium leguminosarum L4 is able to serve as a host for the plasmid RP1. Properties of R. leguminosarum [RP1] plasmid carrier suggest that the expression of RP1-coded Apr gene(s) is inhibited in this host, although the determinants of transfer and resistance to kanamycin and tetracycline are expressed. This system exemplifies a differential expression of plasmid genes in a new host.     

大肠杆菌AroG蛋白F209S突变体对苯丙氨酸反馈抑制作用的影响(英文)

在大肠杆菌中 ,80 %的 3 脱氧 D 阿拉伯 庚酮 7 磷酸 (3 deoxy D arabino heptulosonate 7 phosphate,DAHP)合酶由aroG基因编码。分别以大肠杆菌K1 2及其抗苯丙氨酸类似物的突变体总DNA为模板 ,以PCR方法扩增得到aroG基因及其突变体。基因测序结果表明抗苯丙氨酸类似物的突变体 ,其aroG基因核苷酸 62 5位发生了T→C的点突变 ,从而使AroG蛋白的 2 0 9位氨基酸由Ser取代了Phe。aroG基因及其突变体克隆到表达质粒 pTrc99A上 ,在大肠杆菌JM 1 0 5中进行表达 ,表达产物的SDS PAGE上可以看到分子量相当明显的条带 ;菌体粗提物中DAHP合酶的活性提高了 1 .8倍 ;酶活抗性检测表明AroG蛋白突变体在一定程度上解除了苯丙氨酸的反馈抑制作用 ;与含K1 2aroG基因的JM1 0 5细胞相比 ,含aroG基因突变体的JM1 0 5细胞可以在含高浓度苯丙氨酸类似物的培养基上生长。     

Variations in R-plasmid DNA concentrations of Escherichia coli during starvation in sewage and brackish waters

Cell culturability and plasmid stability in Escherichia coli containing plasmids RP1, R388 and pUB824 were studied in raw and treated wastewater, and in brackish water. The E. coli strain survived well in the three samples of water employed. Moreover, the three plasmids were maintained under all conditions studied. Interestingly, plasmid DNA concentration of individual plasmids followed the same evolution as the culturable bacteria in the corresponding selective medium when the bacteria grew in raw or treated wastewater. In contrast, in brackish water, the stress due to the oligotrophic and salinity conditions of the medium produced an initial paradoxical increase in plasmid DNA concentration, followed by a decrease in the number of culturable bacteria in the corresponding selective medium. Maintenance of RP1 (56 kbp) and R388 (33 kbp) was markedly influenced by nutritive conditions, which caused a segregation of the plasmids from cells. The results of the present study suggest that variations in plasmid DNA concentrations in an aquatic environment depend on the quality of the water and also on the molecular weight of the plasmid considered.     

Resistance to trifluoroperazine,a calmodulin inhibitor,maps to the fabD locus in Escherichia coli

A mutant, tfpA1, resistant to the calmodulin inhibitor trifluoroperazine (TFP) at 30°C, was isolated in Escherichia coli. The mutant showed a reduced growth rate at 30°C and was temperature sensitive (ts) at 42°C for growth, forming short filaments. The mutation was mapped to the 24 min region of the chromosome and the gene was cloned by complementation of the is defect. Subsequent subcloning, complementation analysis, marker rescue mapping and sequencing, identified tfpA as fabD, encoding the 35 kDa, malonyl-coenzyme A transacylase (MCT) enzyme, required for the initial step in the elongation cycle for fatty acid biosynthesis. Resistance to TFP may result from altered permeability of the cell envelope, although the mutant remained sensitive to other calmodulin inhibitors and to other antibacterial agents. Alternatively, resistance may be more indirect, resulting from alterations in intracellular Ca⁺⁺ levels which affect the activity of the TFP target in some way.     

The effect of environmental iron concentration on R-plasmid RP1-mediated changes in the lipopolysaccharide and fatty acid composition of Proteus mirabilis

Abstract Rhizobium meliloti JJ-1 grown in a medium rich in manganese elaborated an exopolysaccharide (EPS) that showed marked differences from that produced in the same medium deficient in added manganese. Glucose, galactose and mannose appeared to be the primary carbohydrate constituents of the EPS isolated from the spent fluid of the control culture, while that from the manganese-enriched broth was comprised mainly of glucose and galactose. Examination of these biopolymers by ¹³C-NMR spectroscopy also revealed substantial differences in their non-carbohydrate moieties.     

Formation of Escherichia coli Hfr strains by integrative suppression with the P group plasmid RP1.

Hfr strains of Escherichia coli were obtained by integrative suppression of a dnaA(Ts) mutation by the Inc P-1 plasmid RP1 without prior creation of an unnatural homology between the plasmid and the E. coli chromosome. Unmodified RP1 mobilized the polarized transfer of the chromosome in a counterclock-wise direction from a distinct origin between 81 min (pyrE) and 82 min (dnaA) with pyrE as a leading marker. Inheritance of RP1-Hfr chromosomal and antibiotic resistance genes was due to recombination with the recipient chromosome, as shown by the need for a functional recA system. The acquisition of temperature resistance and donor ability was accompanied by the disappearance of free plasmid when the selection pressure for integration was maintained (growth at 41 degrees C); the loss of temperature resistance and donor ability was accompanied by the reappearance of autonomous RP1 when the selection pressure was removed (growth at 30 degrees C).