生物
噬菌体
重组酶
细胞生物学
Lac抑制因子
指数增长
遗传学
生物过程
抑制因子
合成生物学
大肠杆菌
基因
紫胶操纵子
基因表达
数学分析
重组
古生物学
数学
作者
Marje Kasari,Villu Kasari,Mirjam Kärmas,Arvi Jõers
标识
DOI:10.1021/acssynbio.1c00618
摘要
Efficient production of biochemicals and proteins in cell factories frequently benefits from a two-stage bioprocess in which growth and production phases are decoupled. Here, we describe a novel growth switch based on the permanent removal of the origin of replication (oriC) from the Escherichia coli chromosome. Without oriC, cells cannot initiate a new round of replication, and they stop growing while their metabolism remains active. Our system relies on a serine recombinase from bacteriophage phiC31 whose expression is controlled by the temperature-sensitive cI857 repressor from phage lambda. The reporter protein expression in switched cells continues after cessation of growth, leading to protein levels up to 5 times higher compared to nonswitching cells. Switching induces a unique physiological state that is different from both normal exponential and stationary phases. The switched cells remain in this state even when not growing, retain their protein synthesis capacity, and do not induce proteins associated with the stationary phase. Our switcher technology is potentially useful for a range of products and applicable in many bacterial species for decoupling growth and production.
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