scholarly journals Synthesis of the N-terminal half of the tetracycline resistance protein of pBR322 is responsible for a high level of plasmid DNA supercoiling in Escherichia coli.

1992 ◽  
Vol 38 (4) ◽  
pp. 379-383
Author(s):  
TETSUYA MURAKAMI ◽  
SATOSHI ISHII ◽  
NAOKI KOMIYAMA ◽  
KAZUO SHISHIDO
Keyword(s):  
Escherichia Coli ◽  
Plasmid Dna ◽  
Tetracycline Resistance ◽  
Dna Supercoiling ◽  
Resistance Protein ◽  
High Level

scholarly journals Plasmid DNA Supercoiling and Gyrase Activity in Escherichia coli Wild-Type and rpoS Stationary-Phase Cells

2003 ◽  
Vol 185 (3) ◽  
pp. 1097-1100 ◽  
Author(s):  
Yazmid Reyes-Domínguez ◽  
Gabriel Contreras-Ferrat ◽  
Jesús Ramírez-Santos ◽  
Jorge Membrillo-Hernández ◽  
M. Carmen Gómez-Eichelmann
Keyword(s):  
Escherichia Coli ◽  
Stationary Phase ◽  
Plasmid Dna ◽  
Bimodal Distribution ◽  
Dna Supercoiling ◽  
Wild Type

ABSTRACT Stationary-phase cells displayed a distribution of relaxed plasmids and had the ability to recover plasmid supercoiling as soon as nutrients became available. Preexisting gyrase molecules in these cells were responsible for this recovery. Stationary-phase rpoS cells showed a bimodal distribution of plasmids and failed to supercoil plasmids after the addition of nutrients, suggesting that rpoS plays a role in the regulation of plasmid topology during the stationary phase.

scholarly journals A Combination of sbmA and tolC Mutations in Escherichia coli K-12 Tn10-Carrying Strains Results in Hypersusceptibility to Tetracycline

2007 ◽  
Vol 190 (4) ◽  
pp. 1491-1494 ◽  
Author(s):  
Ricardo E. de Cristóbal ◽  
Paula A. Vincent ◽  
Raúl A. Salomón
Keyword(s):  
Escherichia Coli ◽  
Double Mutant ◽  
Phenotypic Expression ◽  
Tetracycline Resistance ◽  
Additive Effect ◽  
Complete Suppression ◽  
High Level ◽  
K 12 ◽  
Level Resistance

ABSTRACT Previously, we demonstrated that Escherichia coli tolC mutations reduce the high-level resistance to tetracycline afforded by the transposon Tn10-encoded TetA pump from resistance at 200 μg/ml to resistance at 40 μg/ml. In this study, we found that the addition of an sbmA mutation to a tolC::Tn10 mutant exacerbates this phenotype: the double mutant did not form colonies, even in the presence of tetracycline at a concentration as low as 5 μg/ml. Inactivation of sbmA alone partially inhibited high-level tetracycline resistance, from resistance at 200 μg/ml to resistance at 120 μg/ml. There thus appears to be an additive effect of the mutations, resulting in almost complete suppression of the phenotypic expression of Tn10 tetracycline resistance.

scholarly journals Effects of Ribosomal Protein S10 Flexible Loop Mutations on Tetracycline and Tigecycline Susceptibility of Escherichia coli

2021 ◽  
Vol 12 ◽  
Author(s):  
Norbert Izghirean ◽  
Claudia Waidacher ◽  
Clemens Kittinger ◽  
Miriam Chyba ◽  
Günther Koraimann ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Ribosomal Proteins ◽  
Parent Strain ◽  
Ribosomal Subunit ◽  
Tetracycline Resistance ◽  
E Coli ◽  
Growth Defects ◽  
Flexible Loop ◽  
Cold Sensitive ◽  
High Level

Tigecycline is a tetracycline derivative that is being used as an antibiotic of last resort. Both tigecycline and tetracycline bind to the small (30S) ribosomal subunit and inhibit translation. Target mutations leading to resistance to these antibiotics have been identified both in the 16S ribosomal RNA and in ribosomal proteins S3 and S10 (encoded by the rpsJ gene). Several different mutations in the S10 flexible loop tip residue valine 57 (V57) have been observed in tigecycline-resistant Escherichia coli isolates. However, the role of these mutations in E. coli has not yet been characterized in a defined genetic background. In this study, we chromosomally integrated 10 different rpsJ mutations into E. coli, resulting in different exchanges or a deletion of S10 V57, and investigated the effects of the mutations on growth and tigecycline/tetracycline resistance. While one exchange, V57K, decreased the minimal inhibitory concentration (MIC) (Etest) to tetracycline to 0.75 μg/ml (compared to 2 μg/ml in the parent strain) and hence resulted in hypersensitivity to tetracycline, most exchanges, including the ones reported previously in resistant isolates (V57L, V57D, and V57I) resulted in slightly increased MICs to tigecycline and tetracycline. The strongest increase was observed for the V57L mutant, with a MIC (Etest) to tigecycline of 0.5 μg/ml (compared to 0.125 μg/ml in the parent strain) and a MIC to tetracycline of 4.0 μg/ml. Nevertheless, none of these exchanges increased the MIC to the extent observed in previously described clinical tigecycline-resistant isolates. We conclude that, next to S10 mutations, additional mutations are necessary in order to reach high-level tigecycline resistance in E. coli. In addition, our data reveal that mutants carrying S10 V57 exchanges or deletion display growth defects and, in most cases, also thermosensitivity. The defects are particularly strong in the V57 deletion mutant, which is additionally cold-sensitive. We hypothesize that the S10 loop tip residue is critical for the correct functioning of S10. Both the S10 flexible loop and tigecycline are in contact with helix h31 of the 16S rRNA. We speculate that exchanges or deletion of V57 alter the positioning of h31, thereby influencing both tigecycline binding and S10 function.

Cloning of tetracycline-resistance genes from various strains of Clostridium perfringens and expression in Escherichia coli

1992 ◽  
Vol 38 (3) ◽  
pp. 215-221 ◽  
Author(s):  
Nitin K. Saksena ◽  
Nicole Truffaut
Keyword(s):  
Escherichia Coli ◽  
Gene Cloning ◽  
Key Words ◽  
Clostridium Perfringens ◽  
Resistance Genes ◽  
Tetracycline Resistance ◽  
Tetracycline Resistance Genes ◽  
Expression In Escherichia Coli ◽  
Large Plasmids ◽  
High Level

One hundred strains of Clostridium perfringens and 52 strains of other Clostridia of human and animal origins were screened for tetracycline resistance. Fifty-six strains were resistant to tetracycline in the C. perfringens group. Ten strains were selected for their high level of resistance. In all of them, the tetracycline-resistance genes were found to be residing in large plasmids of about 50 kb, all showing homologies. Several tetracycline-resistance genes from plasmids of various strains of C. perfringens were cloned in plasmid pUC19 and the resistance was expressed in Escherichia coli. Hybridization analysis showed these genes to be homologous among themselves and also to tetP gene from the PCW3-type plasmid. Key words: tetracycline resistance, Clostridium perfringens, gene cloning, recombinant plasmids.

scholarly journals Incidence of Antibiotic Resistance in Campylobacter jejuni Isolated in Alberta, Canada, from 1999 to 2002, with Special Reference to tet(O)-Mediated Tetracycline Resistance

2004 ◽  
Vol 48 (9) ◽  
pp. 3442-3450 ◽  
Author(s):  
Amera Gibreel ◽  
Dobryan M. Tracz ◽  
Lisa Nonaka ◽  
Trinh M. Ngo ◽  
Sean R. Connell ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Campylobacter Jejuni ◽  
Tetracycline Resistance ◽  
Clinical Isolates ◽  
Nucleotide Substitutions ◽  
The Past ◽  
Genes Encoding ◽  
Large Plasmids ◽  
High Level

ABSTRACT Of 203 human clinical isolates of Campylobacter jejuni from Alberta, Canada (1999 to 2002), 101 isolates (50%) were resistant to at least 64 μg of tetracycline/ml, with four isolates exhibiting higher levels of tetracycline resistance (512 μg/ml). In total, the MICs for 37% of tetracycline-resistant isolates (256 to 512 μg/ml) were higher than those previously reported in C. jejuni (64 to 128 μg/ml). In the tetracycline-resistant clinical isolates, 67% contained plasmids and all contained the tet(O) gene. Four isolates resistant to high levels of tetracycline (MIC = 512 μg/ml) contained plasmids carrying the tet(O) gene, which could be transferred to other isolates of C. jejuni. The tetracycline MICs for transconjugants were comparable to those of the donors. Cloning of tet(O) from the four high-level tetracycline-resistant isolates conferred an MIC of 32 μg/ml for Escherichia coli DH5α. In contrast, transfer to a strain of C. jejuni by using mobilization conferred an MIC of 128 μg/ml. DNA sequence analysis determined that the tet(O) genes encoding lower MICs (64 to 128 μg/ml) were identical to one other, although the tet(O) genes encoding a 512-μg/ml MIC demonstrated several nucleotide substitutions. The quinolone resistance determining region of four ciprofloxacin-resistant isolates (2%) was analyzed, and resistance was associated with a chromosomal mutation in the gyrA gene resulting in a Thr-86-Ile substitution. In addition, six kanamycin-resistant isolates contained large plasmids that carry the aphA-3 marker coding for 3′-aminoglycoside phosphotransferase. Resistance to erythromycin was not detected in 203 isolates. In general, resistance to most antibiotics in C. jejuni remains low, except for resistance to tetracycline, which has increased from about 8 to 50% over the past 20 years.

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