Creating Bacterial Strains from Genomes That Have Been Cloned and Engineered in Yeast
Abstract
Character Transplant: When engineering bacteria, it can be advantageous to propagate the genomes in yeast. However, to be truly useful, one must be able to transplant the bacterial chromosome from yeast back into a recipient bacterial cell. But because yeast does not contain restriction-modification systems, such transplantation poses problems not encountered in transplantation from one bacterial cell to another. Bacterial genomes isolated after growth in yeast are likely to be susceptible to the restriction-modification system(s) of the recipient cell, as well as their own. Lartigue et al. (p. 1693 , published online 20 August) describe multiple steps, including in vitro DNA methylation, developed to overcome such barriers. A Mycoplasma mycoides large-colony genome was propagated in yeast as a centromeric plasmid, engineered via yeast genetic systems, and, after specific methylation, transplanted into M. capricolum to produce a bacterial cell with the genotype and phenotype of the altered M. mycoides large-colony genome.
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Genome manipulation in yeast and transplantation do not induce mutations.
“Genome manipulation in yeast and transplantation do not induce mutations as shown by sequencing before and after transplantation (Lartigue et al., 2009)”
From Genetic requirements for cell division in a genomically minimal cell (2020) · cites this paper as Lartigue et al., 2009
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“Genome transplantation Mycoplasmal genomes that are maintained and manipulated in yeast as yeast centromeric plasmids can be booted up by genome transplantation into a recipient organism,Mycoplasma capricolum, as described in detail elsewhere (Gibson et al., 2010; Hutchison et al., 2016; Lartigue et al., 2009; Tsarmpo…”
BackgroundMethodSelf-citation
- Authors
- Lartigue, C., Vashee, S., Algire, M. A., Chuang, R.-Y., Benders, G. A., Ma, L., Noskov, V. N., Denisova, E. A., Gibson, D. G., Assad-Garcia, N., Alperovich, N., Thomas, D. W., Merryman, C., Hutchison, C. A., Smith, H. O., Venter, J. C., & Glass, J. I.
- Affiliations
- J. Craig Venter Institute (US)
- Corresponding author
- Vashee, Sanjay
- Thomas, David W.
- ORCID
- Venue
- Science
- Published
- 2009-09-25
- Volume
- 325
- Issue
- 5948
- Pages
- 1693-1696
- ISSN
- 0036-8075 (ISSN-L)
- 1095-9203
- OpenAlex
- W2134370272
- Cited by
- 332
- References
- 0
- Citations by year
- 2026: 10 · 2025: 13 · 2024: 20 · 2023: 25 · 2022: 15 · 2021: 12 · 2020: 14 · 2019: 17 · 2018: 17 · 2017: 10 · 2016: 12 · 2015: 17 · 2014: 13 · 2013: 22 · 2012: 21
- FWCI
- 15.45
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Science (publishedVersion)
- PubMed (publishedVersion)
- HAL (submittedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Keywords
- Mycoplasma mycoides, Mycoplasma capricolum, restriction-modification systems, DNA methylation, centromeric plasmids, bacterial genome engineering
- MeSH terms
- Centromere, Cloning, Molecular, DNA Restriction Enzymes, Genetic Engineering, Mycoplasma mycoides, Plasmids, Saccharomyces cerevisiae, Transformation, Bacterial, Deoxyribonucleases, Type III Site-Specific, Genome, Bacterial, Sequence Deletion, Sequence Analysis, DNA, Gene Transfer Techniques, DNA Methylation, Mycoplasma capricolum
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