title: Identification and modeling of a novel chloramphenicol resistance protein detected by functional metagenomics in a wetland of Lerma, Mexico.
abstract: The exploration of novel antibiotic resistance determinants in a particular environment may be limited because of the presence of uncultured microorganisms. In this work, a culture-independent approach based on functional metagenomics was applied to search for chloramphenicol resistance genes in agro-industrial wastewater in Lerma de Villada, Mexico. To this end, a metagenomic library was generated in Escherichia coli DH10B containing DNA isolated from environmental samples of the residual arsenic-enriched (10 mg/ml) effluent. One resistant clone was detected in this library and further analyzed. An open reading frame similar to a multidrug resistance protein from Aeromonas salmonicida and responsible for chloramphenicol resistance was identified, sequenced, and found to encode a member of the major facilitator superfamily (MFS). Our results also showed that the expression of this gene restored streptomycin sensitivity in E. coli DH10B cells. To gain further insight into the phenotype of this MFS family member, we developed a model of the membrane protein multiporter that, in addition, may serve as a template for developing new antibiotics.
authors.affiliation: Environmental Sciences Department, Metropolitan Autononous University (Lerma Unit), Lerma de Villada, Mexico.
authors.forename: Marcos
authors.initials: M
authors.lastname: López-Pérez
authors.affiliation: Department of Molecular Evolution, Center of Astrobiology (CSIC-INTA), Torrejón de Ardoz, Spain.
authors.forename: Salvador
authors.initials: S
authors.lastname: Mirete
authors.affiliation: Earth Resources Department, Metropolitan Autononous University (Lerma Unit), Lerma de Villada, Mexico.
authors.forename: Eduardo
authors.initials: E
authors.lastname: Jardón-Valadez
authors.affiliation: Department of Molecular Evolution, Center of Astrobiology (CSIC-INTA), Torrejón de Ardoz, Spain.
authors.forename: José E
authors.initials: JE
authors.lastname: González-Pastor
mesh_terms: D048409:Aeromonas salmonicida / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D002702:Chloramphenicol Resistance; D003001:Cloning, Molecular; D005618:Fresh Water / Q000382:microbiology*; D056186:Metagenomics; D008800:Mexico; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D053833:Wetlands
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins