Isolation of a conjugative F-like plasmid from a multidrug-resistant Escherichia coli strain CM6 using tandem shock wave-mediated transformation

• Published in: Journal of microbiological methods 2015-07, Vol.114, p.1-8
• Main Authors: Soto-Alonso, G., Cruz-Medina, J.A., Caballero-Pérez, J., Arvizu-Hernández, I., Ávalos-Esparza, L.M., Cruz-Hernández, A., Romero-Gómez, S., Rodríguez, A.L., Pastrana-Martínez, X., Fernández, F., Loske, A.M., Campos-Guillén, J.
• Format: Article
• Language: English
• Subjects: Animals; beta-Lactamases - genetics; Cattle; Conjugation, Genetic; DNA, Bacterial - chemistry; DNA, Bacterial - genetics; Drug Resistance, Multiple, Bacterial; Escherichia coli; Escherichia coli - drug effects; Escherichia coli - genetics; Escherichia coli - isolation & purification; F Factor - analysis; F-like plasmid; Feces - microbiology; Gene Transfer Techniques; Multidrug-resistant bacteria; Sequence Analysis, DNA; Sequence Homology; Tandem shock waves; Transformation; Transformation, Bacterial
• ISSN: 0167-7012; 1872-8359
• DOI: 10.1016/j.mimet.2015.04.007

Genetic characterization of plasmids from bacterial strains provides insight about multidrug resistance. Ten wild type Escherichia coli (E. coli) strains isolated from cow fecal samples were characterized by their antibiotic resistance profile, plasmid patterns and three different identification methods. From one of the strains, a fertility factor-like plasmid was replicated using tandem shock wave-mediated transformation. Underwater shock waves with a positive pressure peak of up to approximately 40MPa, followed by a pressure trough of approximately −19MPa were generated using an experimental piezoelectric shock wave source. Three different shock wave energies and a fixed delay of 750μs were used to study the relationship between energy and transformation efficiency (TE), as well as the influence of shock wave energy on the integrity of the plasmid. Our results showed that the mean shock wave-mediated TE and the integrity of the large plasmid (~70kb) were reduced significantly at the energy levels tested. The sequencing analysis of the plasmid revealed a high identity to the pHK17a plasmid, including the replication system, which was similar to the plasmid incompatibility group FII. It also showed that it carried an extended spectrum beta-lactamase gene, ctx-m-14. Furthermore, diverse genes for the conjugative mechanism were identified. Our results may be helpful in improving methodologies for conjugative plasmid transfer and directly selecting the most interesting plasmids from environmental samples.