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. 2023 Aug 31;14:1230723. doi: 10.3389/fpls.2023.1230723

Figure 2.

Figure 2

Functional complementation of the PRK mutant. (A) Design of the pCMM-24 construct used for PRK mutant complementation. (B) PCR on genomic DNA to confirm the presence of the CIB1 insertion in the PRK gene in the ΔPRK strain (Locus PRK mutant) and the presence of the pCMM-24 insertion in the complemented strains (Locus insert) in the complemented strains (Cx strains). (C) Anti-PRK western blot on total soluble protein extracts of the CC-4533, ΔPRK and Cx strains (100% corresponds to 12 µg of total protein). (D) Spot test in TAP and HSM minimal media after 5 days growth at 25°C in the dark or in the light (100 µmol photons m-2 s-1). The number of cells spotted is indicated above the spot test image. To show the growth of the ΔPRK strain in TAP light conditions, the growth after 10 days is shown. (E) Relative quantification of the PRK protein content in the Cx strains compared to CC-4533, calculated from the data in panel (C, F) Total activity of reduced PRK in the different strains. Desalted crude extracts of WT, Cx and ΔPRK were reduced with 20 mM DTT prior to activity measurement. (G) Growth profile of selected strains. Cultures were inoculated at 10s cell s/mL and incubated in TAP under light (25°C and 100 µmol photons m-2 s-1).