Karas Bogumil J, Moreau Nicolette G, Deerinck Thomas J, Gibson Daniel G, Venter J Craig, Smith Hamilton O, Glass John I
Synthetic Biology and Bioenergy Group , J. Craig Venter Institute , La Jolla , California 92037 , United States.
National Centre for Microscopy and Imaging Research , University of California, San Diego , La Jolla , 92093 , United States.
ACS Synth Biol. 2019 Feb 15;8(2):239-244. doi: 10.1021/acssynbio.8b00449. Epub 2019 Jan 24.
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.
我们之前发现，完整的细菌染色体可以直接转移到酵母宿主细胞中，通过将细菌细胞与酿酒酵母原生质球融合，它们能够作为着丝粒质粒进行繁殖。在宿主细胞内，可以对酵母着丝粒质粒引入任意数量的所需基因变化，以产生设计基因组，这些基因组可以通过基因组移植方案得以实现。早期研究表明，从供体细菌（如蕈状支原体、山羊支原体或流感嗜血杆菌）中去除限制系统可提高基因组转移的成功率。这些发现表明，其他遗传因素可能也会影响细菌到酵母的基因组转移过程。在本研究中，我们证明，从蕈状支原体中去除一个特定的遗传因子——甘油摄取促进蛋白基因glpF，可使直接基因组转移显著增加，增幅高达21倍。此外，我们还表明，完整的细菌细胞被酵母原生质球内吞，在这些酵母细胞中产生类似细胞器的结构。这可能会带来创造新型合成细胞器的可能性。
