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Open AccessArticle

Trans-Kingdom Conjugation within Solid Media from Escherichia coli to Saccharomyces cerevisiae

1
Department of Biology, The University of Western Ontario, London, ON N6A 5B7, Canada
2
Designer Microbes Inc., London, ON N5Z 3N2, Canada
3
Department of Biochemistry, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON N6A 5C1, Canada
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Int. J. Mol. Sci. 2019, 20(20), 5212; https://doi.org/10.3390/ijms20205212
Received: 29 September 2019 / Revised: 18 October 2019 / Accepted: 19 October 2019 / Published: 21 October 2019
(This article belongs to the Special Issue Yeast Biology)
Conjugation is a bacterial mechanism for DNA transfer from a donor cell to a wide range of recipients, including both prokaryotic and eukaryotic cells. In contrast to conventional DNA delivery techniques, such as electroporation and chemical transformation, conjugation eliminates the need for DNA extraction, thereby preventing DNA damage during isolation. While most established conjugation protocols allow for DNA transfer in liquid media or on a solid surface, we developed a procedure for conjugation within solid media. Such a protocol may expand conjugation as a tool for DNA transfer to species that require semi-solid or solid media for growth. Conjugation within solid media could also provide a more stable microenvironment in which the conjugative pilus can establish and maintain contact with recipient cells for the successful delivery of plasmid DNA. Furthermore, transfer in solid media may enhance the ability to transfer plasmids and chromosomes greater than 100 kbp. Using our optimized method, plasmids of varying sizes were tested for transfer from Escherichia coli to Saccharomyces cerevisiae. We demonstrated that there was no significant change in conjugation frequency when plasmid size increased from 56.5 to 138.6 kbp in length. Finally, we established an efficient PCR-based synthesis protocol to generate custom conjugative plasmids. View Full-Text
Keywords: conjugation; solid media; Saccharomyces cerevisiae; trans-kingdom; Escherichia coli; pTA-Mob; yeast assembly conjugation; solid media; Saccharomyces cerevisiae; trans-kingdom; Escherichia coli; pTA-Mob; yeast assembly
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Soltysiak, M.P.M.; Meaney, R.S.; Hamadache, S.; Janakirama, P.; Edgell, D.R.; Karas, B.J. Trans-Kingdom Conjugation within Solid Media from Escherichia coli to Saccharomyces cerevisiae. Int. J. Mol. Sci. 2019, 20, 5212.

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