Abstract:
We describe a recombineering-based method for the genetic manipulation of lytically replicating bacteriophages, focusing on mycobacteriophages. The approach utilizes recombineering-proficient strains of Mycobacterium smegmatis and employs a cotransformation strategy with purified phage genomic DNA and a mutagenic substrate, which selects for only those cells that are competent to take up DNA. The cotransformation method, combined with the high rates of recombination obtained in M. smegmatis recombineering strains, allows for the efficient and rapid generation of bacteriophage mutants. © 2019, Springer Science+Business Media, LLC, part of Springer Nature.
Registro:
Documento: |
Artículo
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Título: | Genetic Manipulation of Lytic Bacteriophages with BRED: Bacteriophage Recombineering of Electroporated DNA |
Autor: | Marinelli, L.J.; Piuri, M.; Hatfull, G.F. |
Filiación: | Division of Dermatology, Department of Medicine, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA, United States Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, IQUIBICEN-CONICET, Buenos Aires, Argentina Laboratorio “Bacteriófagos y Aplicaciones Biotecnológicas”, Departamento de Química Biológica, FCEyN, UBA, Ciudad Universitaria, Ciudad Autónoma de Buenos Aires, Argentina Department of Biological Sciences and Pittsburgh Bacteriophage Institute, University of Pittsburgh, Pittsburgh, PA, United States
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Palabras clave: | BRED; Electroporation; Mycobacteria; Mycobacteriophage; Recombineering; bacteriophage DNA; genomic DNA; oligonucleotide; Rac protein; recombinant protein; RecT protein; bacteriophage recombineering of electroporated DNA; bacterium culture; electroporation; genetic engineering; genetic manipulation; genetic recombination; methodology; mutant; mycobacteriophage; Mycobacterium smegmatis; natural science; nonhuman; polymerase chain reaction; prophage; recombineering |
Año: | 2019
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Volumen: | 1898
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Página de inicio: | 69
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Página de fin: | 80
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DOI: |
http://dx.doi.org/10.1007/978-1-4939-8940-9_6 |
Título revista: | Methods in Molecular Biology
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Título revista abreviado: | Methods Mol. Biol.
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ISSN: | 10643745
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Registro: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_10643745_v1898_n_p69_Marinelli |
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Citas:
---------- APA ----------
Marinelli, L.J., Piuri, M. & Hatfull, G.F.
(2019)
. Genetic Manipulation of Lytic Bacteriophages with BRED: Bacteriophage Recombineering of Electroporated DNA. Methods in Molecular Biology, 1898, 69-80.
http://dx.doi.org/10.1007/978-1-4939-8940-9_6---------- CHICAGO ----------
Marinelli, L.J., Piuri, M., Hatfull, G.F.
"Genetic Manipulation of Lytic Bacteriophages with BRED: Bacteriophage Recombineering of Electroporated DNA"
. Methods in Molecular Biology 1898
(2019) : 69-80.
http://dx.doi.org/10.1007/978-1-4939-8940-9_6---------- MLA ----------
Marinelli, L.J., Piuri, M., Hatfull, G.F.
"Genetic Manipulation of Lytic Bacteriophages with BRED: Bacteriophage Recombineering of Electroporated DNA"
. Methods in Molecular Biology, vol. 1898, 2019, pp. 69-80.
http://dx.doi.org/10.1007/978-1-4939-8940-9_6---------- VANCOUVER ----------
Marinelli, L.J., Piuri, M., Hatfull, G.F. Genetic Manipulation of Lytic Bacteriophages with BRED: Bacteriophage Recombineering of Electroporated DNA. Methods Mol. Biol. 2019;1898:69-80.
http://dx.doi.org/10.1007/978-1-4939-8940-9_6