Direct Transfer of a Mycoplasma mycoides Genome to Yeast Is Enhanced by Removal of the Mycoides Glycerol Uptake Factor Gene glpF.
Bogumil J KarasNicolette G MoreauThomas J DeerinckDaniel G GibsonJ Craig VenterHamilton O SmithJohn I GlassPublished in: ACS synthetic biology (2019)
We previously discovered that intact bacterial chromosomes can be directly transferred to a yeast host cell where they can propagate as centromeric plasmids by fusing bacterial cells with S accharomyces cerevisiae spheroplasts. Inside the host any desired number of genetic changes can be introduced into the yeast centromeric plasmid to produce designer genomes that can be brought to life using a genome transplantation protocol. Earlier research demonstrated that the removal of restriction-systems from donor bacteria, such as Mycoplasma mycoides, Mycoplasma capricolum, or Haemophilus influenzae increased successful genome transfers. These findings suggested that other genetic factors might also impact the bacteria-to-yeast genome transfer process. In this study, we demonstrated that the removal of a particular genetic factor, the glycerol uptake facilitator protein gene glpF from M. mycoides, significantly increased direct genome transfer by up to 21-fold. Additionally, we showed that intact bacterial cells were endocytosed by yeast spheroplasts producing organelle-like structures within these yeast cells. These might lead to the possibility of creating novel synthetic organelles.
Keyphrases
- genome wide
- induced apoptosis
- saccharomyces cerevisiae
- cell cycle arrest
- copy number
- dna methylation
- escherichia coli
- cell wall
- randomized controlled trial
- endoplasmic reticulum stress
- cell therapy
- stem cells
- cell death
- signaling pathway
- high resolution
- multidrug resistant
- small molecule
- pi k akt
- bone marrow
- klebsiella pneumoniae