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R Zilhao

Publications and source records attributed to R Zilhao.

6 recordsLinked to original sources

Bacillus subtilis spore coat assembly requires cotH gene expression.

Endospores of Bacillus subtilis are encased in a protein shell, known as the spore coat, composed of a lamella-like inner layer and an electron-dense outer layer. We report the identification and characterization of a gene, herein called cotH, located at 300 degrees on the B. subtilis genetic map between two divergent cot genes, cotB and cotG. The cotH open reading frame extended for 1,086 bp and corresponded to a polypeptide of 42.8 kDa. Spores of a cotH null mutant were normally heat, lysozyme, and chloroform resistant but were impaired in germination. The mutant spores were also pleiotropically deficient in several coat proteins, including the products of the previously cloned cotB, -C, and -G genes. On the basis of the analysis of a cotE cotH double mutant, we infer that CotH is probably localized in the inner coat and is involved in the assembly of several proteins in the outer layer of the coat.

Amino Acid Sequence↗

Nucleotide sequence of the fosB gene conferring fosfomycin resistance in Staphylococcus epidermidis.

We have determined the nucleotide sequence of gene fosB of plasmid pIP1842, which confers resistant to fosfomycin in Staphylococcus epidermidis BM2641. The resistance gene was identified as a coding sequence of 417 bases pairs corresponding to a protein with a calculated Mw of 16,345 daltons. This value is in good agreement with that of 15,000, estimated by SDS-polyacrylamide gel electrophoresis of a bacterial cell-free coupled transcription-translation system. The fosB gene product exhibits 48% sequence homology with the fosfomycin resistance protein FOSA of Enterobacteriaceae. The substitutions are scattered throughout indicating that the two corresponding genes have diverged from a common ancestor. Downstream from fosB is located an open reading frame that was partially sequenced. The deduced amino-acid sequence was 50% homologous to REP alpha 1, a protein involved in the replication of the Enterococcus faecalis plasmid pAM alpha 1.

Amino Acid Sequence↗

Gene heterogeneity for tetracycline resistance in Staphylococcus spp.

Nucleotide sequences related to four tet genes were studied by hybridization in 183 clinical Staphylococcus isolates. tet(K) predominated in strains resistant only to tetracycline, while tet(M) was responsible for combined tetracycline and minocycline resistance. In strains harboring both genes, they contributed additively. tet(L) was detected in only five strains, and no hybridization was observed with tet(O).

DNA, Bacterial↗

Non-radioactive gene probes for the detection of tetracycline and/or minocycline resistance in staphylococci.

This report describes the development of a non-radioactive gene probe for the characterization of tetracycline and/or minocycline resistant staphylococci. Both radio-labelled and non-radioactive gene probes yielded a better characterization of the strains than the determination of minimal inhibitory concentration (MIC). A 100% correlation was found between radioactive and non-radioactive hybridization tests. The latter technique detected genes present in a single copy per bacterial genome, could be performed in 8 h, avoided radioactive hazards and autoradiography delay, and appeared therefore to be a useful tool for clinical and epidemiological studies.

Animals↗

Occurrence of the Campylobacter resistance gene tetO in Enterococcus and Streptococcus spp.

The distribution of nucleotide sequences related to tetK, tetL, tetM, and tetO was studied by dot blot hybridization in 178 strains of Streptococcus and Enterococcus spp. that were resistant to tetracycline. The tetO gene, which is responsible for tetracycline resistance in Campylobacter spp., was detected in six Streptococcus strains and two Enterococcus strains, in which it was borne by similar plasmids. This observation confirms our previous proposal that tetO originated in gram-positive cocci. tetM, the most prevalent resistance gene, was present alone in 109 strains and associated with tetL in 33 strains in which the two genes contributed cooperatively to high-level tetracycline resistance. tetL was present alone in five Enterococcus strains, and tetK was detected in a single Streptococcus strain. The existence of 22 strains that did not hybridize to the probes suggest that tetracycline resistance in streptococci and enterococci involves additional gene classes as well.

Campylobacter↗