Plasmid‐encoded protein attenuates Escherichia coli swimming velocity and cell growth, not reprogrammed regulatory functions

Plasmid‐encoded protein attenuates Escherichia coli swimming velocity and cell growth, not reprogrammed regulatory functions
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质粒编码的蛋白质减弱了大肠杆菌的游动速度和细胞生长,而不是重新编程的调节功能

DOI:
10.1002/btpr.2778
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发表时间:
2019
影响因子:
2.9
通讯作者:
Bentley, William E.
Bentley, William E.
中科院分区:
工程技术4区
文献类型:
--
作者:
Virgile, Chelsea;Hauk, Pricila;Wu, Hsuan‐Chen;Bentley, William E.

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除了设计新的合成途径外,合成生物学家还重新连接细胞以执行“可编程”功能,例如创建伤口愈合益生菌。然而,工程调节电路和合成机器可能对细胞功能有害,特别是如果“代谢负担”很大的话。在这里,一个合成的调控电路,以前构建的直接geophthetics ichiacolito游向过氧化氢,伤口产生的信号,被证明工作,即使与抗生素抗性基因和乳糖利用相关的基因的共表达。然而,我们发现,与组成型表达GFP(作为标记)并另外赋予对卡那霉素和四环素抗性的第二载体共转化导致较慢的速度(Δ~6 μm/s)和显著降低的生长速率(Δ > 50%)。然而,额外的载体并没有改变H2 O2依赖性运动的奔跑翻滚比或方向特征。这种额外负担的主要影响仅限于减缓细胞速度和生长,表明即使在宿主细胞上施加外源性负担时,也可以维持通过最低限度地改变天然调节回路而重编程的细胞运动性。© 2019美国化学工程师学会Biotechnol.程序,35:e2778,2019.
In addition to engineering new pathways for synthesis, synthetic biologists rewire cells to carry out “programmable” functions, an example being the creation of wound‐healing probiotics. Engineering regulatory circuits and synthetic machinery, however, can be deleterious to cell function, particularly if the “metabolic burden” is significant. Here, a synthetic regulatory circuit previously constructed to directEscherichiacolito swim toward hydrogen peroxide, a signal of wound generation, was shown to work even with coexpression of antibiotic resistance genes and genes associated with lactose utilization. We found, however, that cotransformation with a second vector constitutively expressing GFP (as a marker) and additionally conferring resistance to kanamycin and tetracycline resulted in slower velocity (Δ~6 μm/s) and dramatically reduced growth rate (Δ > 50%). The additional vector did not, however, alter the run‐and‐tumble ratio or directional characteristics of H2O2–dependent motility. The main impact of this additional burden was limited to slowing cell velocity and growth, suggesting that reprogrammed cell motility by minimally altering native regulatory circuits can be maintained even when extraneous burden is placed on the host cell. © 2019 American Institute of Chemical EngineersBiotechnol. Prog., 35: e2778, 2019.
运动性对于肿瘤组织中细菌的有效分布和积累至关重要。
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