Isolation of a conjugative F-like plasmid from a multidrug-resistant Escherichia coli strain CM6 using tandem shock wave-mediated transformation.
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 40 MPa, followed by a pressure trough of approximately -19 MPa were generated using an experimental piezoelectric shock wave source. Three different shock wave energies and a fixed delay of 750 μs 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 (~70 kb) 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.
Facultad de Ciencias Naturales, Universidad Autónoma de Querétaro, Avenida de las Ciencias s/n, Querétaro, Qro. 76230, Mexico.
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Soto-Alonso
Facultad de Ciencias Naturales, Universidad Autónoma de Querétaro, Avenida de las Ciencias s/n, Querétaro, Qro. 76230, Mexico.
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Cruz-Medina
Facultad de Ingeniería, Universidad Autónoma de Querétaro, Cerro de las Campanas s/n, Querétaro, Qro. 76010, Mexico.
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Caballero-Pérez
Facultad de Ciencias Naturales, Universidad Autónoma de Querétaro, Avenida de las Ciencias s/n, Querétaro, Qro. 76230, Mexico.
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Arvizu-Hernández
Facultad de Química, Universidad Autónoma de Querétaro, Cerro de las Campanas s/n, Querétaro, Qro. 76010, Mexico.
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Ávalos-Esparza
Facultad de Ingeniería, Universidad Autónoma de Querétaro, Cerro de las Campanas s/n, Querétaro, Qro. 76010, Mexico.
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Cruz-Hernández
Facultad de Química, Universidad Autónoma de Querétaro, Cerro de las Campanas s/n, Querétaro, Qro. 76010, Mexico.
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Romero-Gómez
Centro de Física Aplicada y Tecnología Avanzada, Universidad Nacional Autónoma de México, Boulevard Juriquilla 3001, Querétaro, Qro. 76230, Mexico.
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Rodríguez
Facultad de Ciencias Naturales, Universidad Autónoma de Querétaro, Avenida de las Ciencias s/n, Querétaro, Qro. 76230, Mexico.
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Pastrana-Martínez
Centro de Física Aplicada y Tecnología Avanzada, Universidad Nacional Autónoma de México, Boulevard Juriquilla 3001, Querétaro, Qro. 76230, Mexico.
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Fernández
Centro de Física Aplicada y Tecnología Avanzada, Universidad Nacional Autónoma de México, Boulevard Juriquilla 3001, Querétaro, Qro. 76230, Mexico; División de Ciencias de la Salud, Universidad del Valle de México, Villas del Mesón 1000, Querétaro, Qro. 76230, Mexico. Electronic address: loske@fata.unam.mx.
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Loske
Facultad de Química, Universidad Autónoma de Querétaro, Cerro de las Campanas s/n, Querétaro, Qro. 76010, Mexico. Electronic address: juan.campos@uaq.mx.
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Campos-Guillén
D000818:Animals; D002417:Cattle; D003227:Conjugation, Genetic; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D024901:Drug Resistance, Multiple, Bacterial; D004926:Escherichia coli / Q000187:drug effects* / Q000235:genetics* / Q000302:isolation & purification; D005144:F Factor / Q000032:analysis*; D005243:Feces / Q000382:microbiology; D018014:Gene Transfer Techniques; D017422:Sequence Analysis, DNA; D017385:Sequence Homology; D014169:Transformation, Bacterial; D001618:beta-Lactamases / Q000235:genetics
D004269:DNA, Bacterial; C456080:beta-lactamase CTX-M-14; D001618:beta-Lactamases
Escherichia coli; F-like plasmid; Multidrug-resistant bacteria; Tandem shock waves; Transformation