Abstract
Plasmids are important in carrying antibiotic resistance and other genes between bacterial cells, and a number of methods can be employed to characterize plasmids from clinical isolates. Single colonies typically obtained as part of hospital workflow can undergo S1 nuclease treatment to linearize plasmids followed by pulsed-field gel electrophoresis to enable determination of the number and sizes of plasmids present. Hybridization of S1/PFGE gels can be used to associate replicon types and passenger genes, such as those conferring antibiotic resistance, with a particular plasmid band. Individual plasmids, obtained by conjugation or transformation, can be compared by gel electrophoresis following restriction digestion of plasmid DNA prepared by alkaline lysis methods, including using specialized kits.
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Acknowledgments
CV is supported by grant GNT1107322 and MK was supported by grant GNT1084672, both from the Australian National Health and Medical Research Council.
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Tagg, K.A., Venturini, C., Kamruzzaman, M., Ginn, A.N., Partridge, S.R. (2020). Plasmid DNA Isolation and Visualization: Isolation and Characterization of Plasmids from Clinical Samples. In: de la Cruz, F. (eds) Horizontal Gene Transfer. Methods in Molecular Biology, vol 2075. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9877-7_1
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DOI: https://doi.org/10.1007/978-1-4939-9877-7_1
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