Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:2.3.1.28 (chloramphenicol acetyltransferase)
 5,100 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Little is known about the genetics of Haemophilus ducreyi, the etiologic agent of chancroid. To develop a method for constructing isogenic mutants of this organism that could be utilized in pathogenesis-related studies, electroporation techniques were evaluated as a means of introducing DNA into this organism. Electroporation of the plasmid shuttle vector pLS88 into H. ducreyi yielded approximately 10(6) antibiotic-resistant transformants per microgram of plasmid DNA. Studies of the feasibility of moving mutated genes into H. ducreyi were initiated by using NotI linker insertion and mini-Tn10kan mutagenesis techniques to introduce insertion mutations into cloned H. ducreyi genes encoding cell envelope antigens. In the former case, a gene encoding chloramphenicol acetyltransferase was then inserted into the NotI linker site created in the cloned H. ducreyi gene. The recombinant Escherichia coli strains containing these mutated plasmids no longer expressed the homologous H. ducreyi cell envelope antigens, as evidenced by their lack of reactivity with monoclonal antibody probes for these H. ducreyi proteins. Subsequent electroporation of both circular and linearized forms of plasmids carrying these mutated H. ducreyi genes into the homologous wild-type strain of H. ducreyi yielded antibiotic-resistant transformants which also lacked reactivity with the cell envelope antigen-specific monoclonal antibodies. Southern blot analysis confirmed that homologous recombination had occurred in these monoclonal antibody-unreactive transformants, resulting in the replacement of the wild-type allele with the mutated allele. Allelic exchange was most efficient when linear DNA molecules were used for electroporation. These results indicate that electroporation methods can be utilized to construct isogenic mutants of H. ducreyi.
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PMID:Use of electroporation to construct isogenic mutants of Haemophilus ducreyi. 164 71

Three clinical isolates of Haemophilus ducreyi, representing at least two subtypes, were shown to be resistant to streptomycin and kanamycin. They also produced a beta-lactamase and chloramphenicol acetyltransferase and were resistant to tetracycline. In the three strains the resistance to both aminoglycoside antibiotics was encoded by a plasmid of ca. 4.7 kilobases which apparently did not carry ampicillin, chloramphenicol, or tetracycline resistance genes, as determined after transfer to Escherichia coli by transformation. Resistance to streptomycin and kanamycin was due to the presence of two aminoglycoside phosphotransferases (APH). The enzyme modifying kanamycin was a 3',5"-APH of type I [APH(3',5")-I], as inferred from its substrate profile and immunological cross-reactivity with the APH(3',5")-I encoded by the transposable element Tn903. However, the APH(3',5")-I gene in H. ducreyi did not appear to be carried by Tn903.
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PMID:Plasmid-mediated aminoglycoside phosphotransferases in Haemophilus ducreyi. 301 Aug 43

We examined chloramphenicol-resistant Haemophilus parainfluenzae and Haemophilus ducreyi strains isolated in various parts of the world. The antibiotic resistance determinants were located on conjugative plasmids in H. ducreyi, but were chromosomally located in H. parainfluenzae. Both species produced chloramphenicol acetyltransferases (CATs) that were sensitive to 5,5'-dithiobis(2-nitrobenzoic acid) like the enteric type II and Haemophilus influenzae CAT enzymes, but differed from these enzymes in elution patterns and subunit molecular weight. Southern blot analysis showed the H. parainfluenzae and H. ducreyi CAT genes were molecularly related to the enteric type II class as well as the H. influenzae CAT. Heterogeneity of the physiochemical properties of the CATs was observed; however, the data suggested that all three Haemophilus spp. have a common ancestral source for the CATs.
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PMID:Molecular characterization of chloramphenicol-resistant Haemophilus parainfluenzae and Haemophilus ducreyi. 387 58

The minimal inhibitory concentration (MIC) of chloramphenicol for a clinical isolate of Haemophilus ducreyi, strain CEB-10, was 16 micrograms/ml. This strain was also resistant to tetracycline (MIC = 64 micrograms/ml) and ampicillin. The presence of a chloramphenicol acetyltransferase (CAT) activity was demonstrated.
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PMID:Detection of chloramphenicol acetyltransferase (CAT) activity in a strain of Haemophilus ducreyi. 698 23