Superior Conjugative Plasmids Delivered by Bacteria to Diverse Fungi.

Autor:	Cochrane RR; Department of Biochemistry, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON, Canada, N6A 5C1., Shrestha A; Department of Biochemistry, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON, Canada, N6A 5C1., Severo de Almeida MM; Department of Biochemistry, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON, Canada, N6A 5C1., Agyare-Tabbi M; Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON, Canada, N1G 2W1., Brumwell SL; Department of Biochemistry, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON, Canada, N6A 5C1., Hamadache S; Department of Biochemistry, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON, Canada, N6A 5C1., Meaney JS; Department of Biochemistry, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON, Canada, N6A 5C1., Nucifora DP; Department of Biochemistry, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON, Canada, N6A 5C1., Say HH; Department of Biochemistry, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON, Canada, N6A 5C1., Sharma J; Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON, Canada, N1G 2W1., Soltysiak MPM; Department of Biology, The University of Western Ontario, London, Ontario, Canada, N6A 5B7., Tong C; Department of Biochemistry, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON, Canada, N6A 5C1., Van Belois K; Department of Biology, The University of Western Ontario, London, Ontario, Canada, N6A 5B7., Walker EJL; Department of Biochemistry, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON, Canada, N6A 5C1., Lachance MA; Department of Biology, The University of Western Ontario, London, Ontario, Canada, N6A 5B7., Gloor GB; Department of Biochemistry, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON, Canada, N6A 5C1., Edgell DR; Department of Biochemistry, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON, Canada, N6A 5C1., Shapiro RS; Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON, Canada, N1G 2W1., Karas BJ; Department of Biochemistry, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON, Canada, N6A 5C1.
Jazyk:	angličtina
Zdroj:	Biodesign research [Biodes Res] 2022 Aug 19; Vol. 2022, pp. 9802168. Date of Electronic Publication: 2022 Aug 19 (Print Publication: 2022).
DOI:	10.34133/2022/9802168
Abstrakt:	Fungi are nature's recyclers, allowing for ecological nutrient cycling and, in turn, the continuation of life on Earth. Some fungi inhabit the human microbiome where they can provide health benefits, while others are opportunistic pathogens that can cause disease. Yeasts, members of the fungal kingdom, have been domesticated by humans for the production of beer, bread, and, recently, medicine and chemicals. Still, the great untapped potential exists within the diverse fungal kingdom. However, many yeasts are intractable, preventing their use in biotechnology or in the development of novel treatments for pathogenic fungi. Therefore, as a first step for the domestication of new fungi, an efficient DNA delivery method needs to be developed. Here, we report the creation of superior conjugative plasmids and demonstrate their transfer via conjugation from bacteria to 7 diverse yeast species including the emerging pathogen Candida auris . To create our superior plasmids, derivatives of the 57 kb conjugative plasmid pTA-Mob 2.0 were built using designed gene deletions and insertions, as well as some unintentional mutations. Specifically, a cluster mutation in the promoter of the conjugative gene traJ had the most significant effect on improving conjugation to yeasts. In addition, we created Golden Gate assembly-compatible plasmid derivatives that allow for the generation of custom plasmids to enable the rapid insertion of designer genetic cassettes. Finally, we demonstrated that designer conjugative plasmids harboring engineered restriction endonucleases can be used as a novel antifungal agent, with important applications for the development of next-generation antifungal therapeutics. Competing Interests: The authors declare no competing financial interests. (Copyright © 2022 Ryan R. Cochrane et al.)
Databáze:	MEDLINE
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