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

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.

2015 ◽  
Vol 81 (16) ◽  
pp. 5560-5566 ◽  
Author(s):  
Seung Won Shin ◽  
Min Kyoung Shin ◽  
Myunghwan Jung ◽  
Kuastros Mekonnen Belaynehe ◽  
Han Sang Yoo

ABSTRACTThe aim of this study was to investigate the prevalence and transferability of resistance in tetracycline-resistantEscherichia coliisolates recovered from beef cattle in South Korea. A total of 155E. coliisolates were collected from feces in South Korea, and 146 were confirmed to be resistant to tetracycline. The tetracycline resistance genetet(A) (46.5%) was the most prevalent, followed bytet(B) (45.1%) andtet(C) (5.8%). Strains carryingtet(A) plustet(B) andtet(B) plustet(C) were detected in two isolates each. In terms of phylogenetic grouping, 101 (65.2%) isolates were classified as phylogenetic group B1, followed in decreasing order by D (17.4%), A (14.2%), and B2 (3.2%). Ninety-one (62.3%) isolates were determined to be multidrug resistant by the disk diffusion method. MIC testing using the principal tetracyclines, namely, tetracycline, chlortetracycline, oxytetracycline, doxycycline, and minocycline, revealed that isolates carryingtet(B) had higher MIC values than isolates carryingtet(A). Conjugation assays showed that 121 (82.9%) isolates could transfer a tetracycline resistance gene to a recipient via the IncFIB replicon (65.1%). This study suggests that the high prevalence of tetracycline-resistantE. coliisolates in beef cattle is due to the transferability of tetracycline resistance genes betweenE. colipopulations which have survived the selective pressure caused by the use of antimicrobial agents.


2013 ◽  
Vol 62 (6) ◽  
pp. 851-858 ◽  
Author(s):  
Gong-Zheng Hu ◽  
Yu-Shan Pan ◽  
Hua Wu ◽  
Han Hu ◽  
Rui Xu ◽  
...  

Tetracycline resistance is one of the most frequently encountered resistance properties in bacteria of animal origin. The aim of the present study was to investigate the prevalence and diversity of tetracycline resistance (tet) genes among Escherichia coli clinical isolates from diseased ducks in China and to report the identification and sequencing of the tet(M) gene. The susceptibility of 85 Escherichia coli strains to tetracyclines was determined by broth microdilution, and the presence of tet genes was investigated by multiplex PCR. All of the 85 isolates were fully resistant to both oxytetracycline and tetracycline, and 76.5 % were resistant to doxycycline. Seventy-seven of the isolates (90.6 %) encoded multiple tet genes, with 17.6, 38.8 and 34.1 % encoding two, three and four tet genes, respectively, and only 7.1 % encoded a single tet(A) gene. The MICs of oxytetracycline and tetracycline for all isolates ranged from 16 to ≥128 µg ml−1 with a MIC90 of >128 µg ml−1, regardless of the type or number of tet genes encoded. Isolates containing tet(M) commonly had more than one tet gene per strain. The doxycycline resistance rate in the tet(M)-positive isolates was significantly higher than in the tet(M)-negative isolates (P<0.05). A full-length tet(M) gene, including the promoter region, was obtained by PCR in seven of the 41 tet(M)-positive isolates and was sequenced and cloned. The cloned tet(M) gene conferred resistance to tetracyclines in the recombinant Escherichia coli host strain. These results revealed that, in these isolates, the prevalence of multiple tet genes was strikingly high and that tet(M) played a role in doxycycline resistance.


2004 ◽  
Vol 70 (4) ◽  
pp. 2503-2507 ◽  
Author(s):  
Andrew Bryan ◽  
Nir Shapir ◽  
Michael J. Sadowsky

ABSTRACT Nonselected and natural populations of Escherichia coli from 12 animal sources and humans were examined for the presence and types of 14 tetracycline resistance determinants. Of 1,263 unique E. coli isolates from humans, pigs, chickens, turkeys, sheep, cows, goats, cats, dogs, horses, geese, ducks, and deer, 31% were highly resistant to tetracycline. More than 78, 47, and 41% of the E. coli isolates from pigs, chickens, and turkeys were resistant or highly resistant to tetracycline, respectively. Tetracycline MICs for 61, 29, and 29% of E. coli isolates from pig, chickens, and turkeys, respectively, were ≥233 μg/ml. Muliplex PCR analyses indicated that 97% of these strains contained at least 1 of 14 tetracycline resistance genes [tetA, tetB, tetC, tetD, tetE, tetG, tetK, tetL, tetM, tetO, tetS, tetA(P), tetQ, and tetX] examined. While the most common genes found in these isolates were tetB (63%) and tetA (35%), tetC, tetD, and tetM were also found. E. coli isolates from pigs and chickens were the only strains to have tetM. To our knowledge, this represents the first report of tetM in E. coli.


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