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. 2020 Oct 22;11:587159. doi: 10.3389/fmicb.2020.587159

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Copyright © 2020 Gao, Ma, Qin, Qiu, Zhang, Li, Lou, Diao, Zhao, Shi, Zhang and Kan.

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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Fur regulates the expression of vieSAB, cdgD, vpsA, and vpsU. Bacterial cells were harvested at an OD₆₀₀ value of approximately 0.6. Statistical differences between wild type (WT) and Δfur (^∗ at P < 0.01) were determined by a two-way ANOVA with Tukey’s post hoc. (A) Quantitative PCR (qPCR) assay was employed to determine relative mRNA levels for each target gene in Δfur and WT using a standard curve of recA (reference gene) expression for each RNA preparation. (B) For the luminescence assay, the entire promoter DNA region of each target gene was cloned into a pBBRlux vector, and then introduced into Δfur and WT, to determine the luminescence activity for each strain using an Infinite^® 200 Pro NanoQuant. The lux activity (RLU) was calculated as light units/OD₆₀₀. The minus and positive numbers represented the nucleotide positions upstream and downstream of the translation start site of each target gene.