Cloning and Transplantation of the Mesoplasma florum Genome
| Autor: | Carole Lartigue, Dominick Matteau, Sébastien Rodrigue, Joëlle Brodeur, Vincent Baby, Géraldine Gourgues, Fabien Labroussaa |
|---|---|
| Přispěvatelé: | Département de biologie [Sherbrooke] (UdeS), Faculté des sciences [Sherbrooke] (UdeS), Université de Sherbrooke (UdeS)-Université de Sherbrooke (UdeS), Biologie du fruit et pathologie (BFP), Université Bordeaux Segalen - Bordeaux 2-Institut National de la Recherche Agronomique (INRA)-Université Sciences et Technologies - Bordeaux 1 |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Genome evolution Saccharomyces cerevisiae Proteins Biomedical Engineering Mesoplasma florum Saccharomyces cerevisiae Bacterial genome size Biochemistry Genetics and Molecular Biology (miscellaneous) Genome Mycoplasma capricolum 03 medical and health sciences Bacterial Proteins genome transplantation Cloning Molecular Deoxyribonucleases Type II Site-Specific Gene Hydro-Lyases Cloning Genetics biology [SDV.BA]Life Sciences [q-bio]/Animal biology Gene Expression Profiling whole'genome cloning Gene Transfer Techniques High-Throughput Nucleotide Sequencing Sequence Analysis DNA General Medicine biology.organism_classification Transplantation Restriction enzyme [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology 030104 developmental biology Mutation Entomoplasmataceae Genome Bacterial Plasmids |
| Zdroj: | ACS Synthetic Biology ACS Synthetic Biology, American Chemical Society, 2018, 7 (1), pp.209-217. ⟨10.1021/acssynbio.7b00279⟩ |
| ISSN: | 2161-5063 |
| DOI: | 10.1021/acssynbio.7b00279 |
| Popis: | UMR BFP - Equipe Mollicutes; International audience; Cloning and transplantation of bacterial genomes is a powerful method for the creation of engineered microorganisms. However, much remains to be understood about the molecular mechanisms and limitations of this approach. We report the whole-genome cloning of Mesoplasma florum in Saccharomyces cerevisiae, and use this model to investigate the impact of a bacterial chromosome in yeast cells. Our results indicate that the cloned M. florum genome is subjected to weak transcriptional activity, and causes no significant impact on yeast growth. We also report that the M. florum genome can be transplanted into Mycoplasma capricolum without any negative impact from the putative restriction enzyme encoding gene mfl307. Using whole-genome sequencing, we observed that a small number of mutations appeared in all M. florum transplants. Mutations also arose, albeit at a lower frequency, when the M. capricolum genome was transplanted into M. capricolum recipient cells. These observations suggest that genome transplantation is mutagenic, and that this phenomenon is magnified by increased phylogenetic distance between the genome donor and the recipient cell. No difference in efficiency was detected after three successive rounds of genome transplantation, suggesting that the observed mutations were not selected during the procedure. Taken together, our results provide a more accurate picture of the events taking place during bacterial genome cloning and transplantation. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|