Fig. 6.

Genetic complementation of the RNase III triple mutant alleviates growth defect

The native A0061 gene was targeted to the glpK pseudogene as a neutral site in the Δ0061/2542/0384 genetic background to generate the Δ0061+/2542/0384 strain. (a) Full segregation of the A0061 gene in the glpK locus was confirmed by PCR using flanking (397+398) and gene specific primers (397+399). WT cells were used as the negative size control and the plasmid used in the transformation was used as the positive control (pJCC256). (b) Overnight cultures grown with bubbling in air (38°C, 200 μmol photons m⁻² s⁻¹) were suspended to an OD 730 nm = 0.5 (1 cm cuvette). Tenfold serial dilutions were plated in quadruplicate on media A+ (7.5 μl/spot) and grown for 48 h (120 μmol photons m⁻² s⁻¹) prior to being photographed. A representative image is shown. (c) For liquid growth, strains were diluted to an OD 730 nm = 0.01 (1 cm cuvette) and grown in test tubes bubbled for 12 h at a light intensity of 200 μmol photons m⁻² s⁻¹. Error bars represent s.d. for three biological replicates.