The Structure of the Chloramphenicol Resistance Gene on a Transferable R Plasmid from the Fish Pathogen, Pasteurella piscicida

Abstract

The chloramphenicol resistance gene (pp-cat) was cloned from a transferable R plasmid of Pasteurella piscicida, pSP9351, and the sequence of the gene was determined. Subcloning and deletion analysis localized the resistance gene, pp-cat, to within a 2.3 kb Hinc11-BamH1 fragment. The fragment as a probe hybridized with the type I chloramphenicol acetyltransferase (CAT) gene and did not hybridize to CAT types II, III, and CAT-VA. The fragment hybridized to transferable R plasmids encoded with resistance to chloramphenicol, which were detected from P. piscicida isolated in different years. Nucleotide sequences of the coding and flanking regions of pp-cat (2031 bp) identified an open reading frame coding type I CAT of a molecular mass of about 25,000 Da. Comparison analysis of the sequences outside the cat open reading frame showed also that pp-cat has homology, in part, with the gene that coding for the endonuclease EcoRII and those that flank the cat gene derived from the Acinetobacter baumannii chromosome.

Cloning and nucleotide sequence analysis of the chloramphenicol resistance gene on conjugative R plasmids from the fish pathogenPhotobacterium damselaesubsp.piscicida

“…The Cm r determinants on transferable R plasmids are classified as Type I variant for Photobacterium damselae subsp. piscicida, Type II variants for Edwardsiella tarda and Aeromonas salmonicida, and Type II (isolated after 1980) and the other variants (isolated before 1977) for Vibrio anguillarum (Aoki 1988, 1989, Zhao & Aoki 1992, Kim & Aoki 1993b. However, in this study Cm r determinants on transferable R plasmids were classified as Type II variants for P. damselae subsp.…”

“…Cm r determinants on transferable R plasmids are classified as Type I variant for Photobacterium damselae subsp. piscicida that was isolated in 1993 from Ehime prefecture in Japan (Kim & Aoki 1993b) and as Type II variant for the same bacteria that was isolated in 1985 and 1991 from Nagasaki and Kumamoto in Japan (this study). Moreover, CAT variants encoded on transferable R plasmids in other fish pathogenic bacteria have been different depending upon the bacterial genera and the year of the bacterial isolation as described above.…”

“…A CAT variant of Photobacterium damselae subsp. piscicida was classified as Type I (Kim & Aoki 1993b). In this study, Cm r determinants on transferable R plasmids from P. damselae subsp.…”

Molecular diversity of Photobacterium damselae ssp. piscicida from cultured amberjacks (Seriola spp.) in Japan by pulsed-field gel electrophoresis and plasmid profiles

“…In the previous reports of P. damselae ssp. piscicida , resistance genes derived from transferable R plasmids were reported against the chloramphenicol (Kim and Aoki 1993b; Morii et al. 2003), sulfonamide (Kim and Aoki 1996), tetracycline and kanamycin (Kim and Aoki 1994).…”

Complete DNA Sequence and Analysis of the Transferable Multiple-Drug Resistance Plasmids (R Plasmids) from Photobacterium damselae subsp. piscicida Isolates Collected in Japan and the United States

“…piscicida, the Km r , Tc r , and Cp r genes were flanked by IS26, making them class I transposons, while the Sa r and Tmp r genes were bordered by transposase genes, suggesting that they are class II transposons. The genetic element of a similar, previously described plasmid, pSP9351 (with the structure IS26-Km r gene-IS26-Tc r gene-IS26-Sa r gene-IS26), is regarded as a complex transposon and exhibits a functional hybrid promoter made up of the transposable element IS26 (6,8,9). Direct evidence shows that IS26 provides part of the promoter of the antibiotic resistance operon (IAB) of pBWH77 and has an active role in the evolution and mobility of antibiotic resistance genes (10).…”