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. 2020 Apr 18;23(5):101067. doi: 10.1016/j.isci.2020.101067

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© 2020 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

PMC Copyright notice

Verification of the Function of the Heme Dynamic Regulation System

(A) Schematic diagram of dynamic control system acting on both hemB and mkate2.

(B) Red fluorescence intensity of strains containing both sgRNA targeting mkate2 and hemB. SH0 (mkate2 is not inhibited by CRISPRi), SH1 (sgRNA-A and sgRNA-1), SH2 (sgRNA-A and sgRNA-2), SH3 (sgRNA-A and sgRNA-3), SOH2 (removed hrtO based on SH2, making CRISPRi constitutive expression), SB4 (only contains sgRNA-A, no sgRNA targeting hemB).

(C) Red fluorescence intensity change after replacing the mkate2 promoter and RBS, SH2-AL: Ptac and RBS_B0034; SH2-AL-1: P_J23110 and RBS_B0032; SH2-AL-2: P_J23101 and RBS_B0032; SH2-AL-3: P_J23106 and RBS_B0032.

(D) The relative expression level of dcas9 in SH2-AL changes with time.

(E) The relative expression level of hemB in SH2-AL changes with time. Error bars represent ±1 SD from the mean of three replicate cultures.