Extended Data Fig. 10: Selection of the Rubisco small subunit mutant strains for phenotype analysis.
From: The structural basis of Rubisco phase separation in the pyrenoid
a, The Rubisco small subunit-less mutant T60 (Δrbcs) was transformed with DNA encoding wild-type and mutant Rubisco small subunits (RBCS) to produce candidate transformants with the genotypes Δrbcs;RBCSWT, Δrbcs;RBCSD23A/E24A, and Δrbcs;RBCSM87D/V94D. Total protein extracts for three strains from each transformation were separated on a polyacrylamide gel. b, The gel shown in panel a was probed by Western blot using a polyclonal antibody mixture that detects both large and small Rubisco subunits. The experiments shown in panel a and b were performed once for selecting the candidate transformants with the highest RBCS expression level from each genotype, in case any phenotype may be caused by low expression level of Rubisco. Selected strains are indicated by an arrow below the lanes and were used for the subsequent phenotypic analyses shown in Fig. 5 and panel c. c, Additional representative TEM images of whole cells of the strains expressing wild-type, D23A/E24A, and M87D/V94D Rubisco small subunit. Scale bar = 500 nm. For each strain, at least 25 images (one image for one cell) were taken and showing similar results.
