Abstract：
:Mycoplasma hominis is a minimal human pathogen that is responsible for genital and neonatal infections. Despite many attempts, there is no efficient genetic tool to manipulate this bacterium, limiting most investigations of its pathogenicity and its uncommon energy metabolism that relies on arginine. The recent cloning and subsequent engineering of other mycoplasma genomes in yeast opens new possibilities for studies of the genomes of genetically intractable organisms. Here, we report the successful one-step cloning of the M. hominis PG21 genome in yeast using the transformation-associated recombination (TAR) cloning method. At low passages, the M. hominis genome cloned into yeast displayed a conserved size. However, after ∼60 generations in selective media, this stability was affected, and large degradation events were detected, raising questions regarding the stability of large heterologous DNA molecules cloned in yeast and the need to minimize host propagation. Taking these results into account, we selected early passage yeast clones and successfully modified the M. hominis PG21 genome using the CRISPR/Cas9 editing tool, available in Saccharomyces cerevisiae. Complete M. hominis PG21 genomes lacking the adhesion-related vaa gene were efficiently obtained.
journal_name
ACS Synth Bioljournal_title
ACS synthetic biologyauthors
Rideau F,Le Roy C,Descamps ECT,Renaudin H,Lartigue C,Bébéar Cdoi
10.1021/acssynbio.6b00379subject
Has Abstractpub_date
2017-05-19 00:00:00pages
891-901issue
5issn
2161-5063journal_volume
6pub_type
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journal_title：ACS synthetic biology
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journal_title：ACS synthetic biology
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