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. 2019 Feb 20;10:278. doi: 10.3389/fmicb.2019.00278

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Copyright © 2019 Qin, Zhao, Yu, Liu, Chu, Tian and Wu.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Flowchart of the genome replication engineering-assisted continuous evolution (GREACE) method. To improve cadmium resistance, pQ-1, pQ-2, and pQ-3 were transformed into host cells [BL21(DE3)] and cultivated under conditions with gradually increasing selective strength. Evolved strains with higher cadmium resistance were selected, and the plasmids were sequenced for confirmation. Genetically modified pQ mutants were eliminated from the evolved strains to stabilize the obtained genotypes and phenotypes.