Bifidobacterium transformation

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Date
2021-03-03
Authors
Hoedt, Emily C.
Bongers, Roger S.
Bottacini, Francesca
Knol, Jan
MacSharry, John
van Sinderen, Douwe
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Springer Science+Business Media, LLC
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Abstract
The protocol presented in this chapter describes a generic method for electrotransformation of Bifidobacterium spp., outlining a technique that is ideal for conferring selective properties onto strains as well as allowing the user to introduce or knock out/in selected genes for phenotypic characterization purposes. We have generalized on the plasmid chosen for transformation and antibiotic selection marker, but the protocol is versatile in this respect and we are able to achieve transformation efficiencies up to 107 transformants/μg of DNA.
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Keywords
Bifidobacterial , Electroporation , Genetic accessibility , Probiotic
Citation
Hoedt, Emily C.; Bongers, Roger S.; Bottacini, Francesca; Knol, Jan; MacSharry, John; van Sinderen, Douwe (2021) 'Bifidobacterium transformation', in van Sinderen, D. and Ventura, M. (eds) Bifidobacteria - Methods and Protocols. Methods in Molecular Biology, vol 2278, pp. 13-19. New York, NY: Humana Press. doi: 10.1007/978-1-0716-1274-3_2
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© 2021, Springer Science+Business Media, LLC, part of Springer Nature. This is a post-peer-review, pre-copyedit version of a chapter published in van Sinderen, D. and Ventura, M. (eds) Bifidobacteria - Methods and Protocols. Methods in Molecular Biology, vol 2278, pp. 13-19. New York, NY: Humana Press. The final authenticated version is available online at: https://doi.org/10.1007/978-1-0716-1274-3_2