Akeno Yuya, Ying Bei-Wen, Tsuru Saburo, Yomo Tetsuya
Graduate School of Information Science and Technology, Osaka University, 1-5 Yamadaoka, Suita, Osaka 565-0871, Japan.
Microb Cell Fact. 2014 Apr 1;13(1):49. doi: 10.1186/1475-2859-13-49.
Host-plasmid interactions have been discussed largely in terms of the influences of plasmids, whereas the contributions of variations in host genomes to host interactions with foreign DNA remain unclear. A strain with a so-called "clean genome" (i.e., MDS42) of reduced genome size has recently been generated from the wild-type strain MG1655, a commonly used host strain. A quantitative evaluation of the influence of plasmid burdens in these two Escherichia coli strains can not only provide an understanding of how a reduced genome responds to foreign DNA but also offer insights into the proper application of these strains.
The decreases in growth caused by the cost of carrying foreign DNA were similar for the wild-type and clean-genome strains. A negative correlation between the growth rate and the total amount of exogenous DNA was observed in both strains, but a better theoretical fit with a higher statistical significance was found for the strain with the clean genome. Compared to the wild-type strain, the clean-genome strain exhibited a reduced carrying capacity for exogenous DNA, which was largely attributed to its ability to restrict the replication of foreign DNA. A tendency to allocate energy and resources toward gene expression, but not DNA replication, was observed in the strain with the clean genome.
The possession of a clean genome constrained the plasmid copy number to a wild-type-equivalent load. The results indicate that the wild-type strain possesses a greater tolerance for foreign DNA, as in endosymbiosis, and that the use of strains with clean genomes will be favorable in the applications that require precise control and theoretical prediction.
宿主 - 质粒相互作用在很大程度上是从质粒的影响方面进行讨论的,而宿主基因组变异对外源DNA相互作用的贡献仍不清楚。最近从常用宿主菌株野生型MG1655中构建出了一种基因组大小减小的所谓“干净基因组”菌株(即MDS42)。对这两种大肠杆菌菌株中质粒负担影响的定量评估,不仅能让我们了解基因组减小后对外源DNA的反应,还能为这些菌株的合理应用提供见解。
携带外源DNA的成本导致的野生型和干净基因组菌株生长下降情况相似。在两种菌株中均观察到生长速率与外源DNA总量之间呈负相关,但干净基因组菌株的理论拟合更好且具有更高的统计学显著性。与野生型菌株相比,干净基因组菌株对外源DNA的携带能力降低,这主要归因于其限制外源DNA复制的能力。在干净基因组菌株中观察到一种将能量和资源分配用于基因表达而非DNA复制的趋势。
拥有干净基因组将质粒拷贝数限制在与野生型相当的水平。结果表明,野生型菌株对外源DNA具有更大的耐受性,如同内共生现象,并且在需要精确控制和理论预测的应用中,使用干净基因组菌株将更具优势。