CC-5034 rbcL-AG265/rbcS1-SSAT mt+ (AG265/SSAT)
$30.00
From Robert J. Spreitzer, University of Nebraska, November 2014
Phenotype: requires elevated CO2 for photosynthetic growth
Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆/rbcS1-SSAT pf2 mt+ (CC-4927), Boon Hoe Lim in Spreitzer’s group created 12 substitutions (V149Q, G168P, V221C, C256F, K258R, I265V, I282H, L326I, M349L, M375L, A398S, and C399V) together in the Rubisco large subunit. This mutant (also named AG265/SSAT) represents one of 15 “associated groups” of amino acids that differ between Chlamydomonas and land plants (Du et al. 2003). It was created to investigate phylogenetic differences that influence Rubisco catalysis (Spreitzer et al. 2005). The 12 substitutions in the large subunit must be complemented by the presence of the Arabidopsis small subunit (SSAT) to produce a functional Rubisco holoenzyme (Lim and Spreitzer, unpublished). The mutant strain requires 5% CO2 in air for photosynthetic growth (Genkov et al. 2010), and has a decrease in Rubisco holoenzyme stability (Lim and Spreitzer, unpublished). It has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.
Du YC, Peddi SR, Spreitzer RJ (2003) Assessment of structural and functional divergence far from the large subunit active site of ribulose-1,5-bisphosphate carboxylase/oxygenase. J Biol Chem 278:49401-49405
Genkov T, Meyer M, Griffiths H, Spreitzer RJ (2010) Functional hybrid Rubisco enzymes with plant small subunits and algal large subunits: Engineered rbcS cDNA for expression in Chlamydomonas. J Biol Chem 285:19833-19841
Spreitzer RJ, Peddi SR, Satagopan S (2005) Phylogenetic engineering at an interface between large and small subunits imparts land-plant kinetic properties to algal Rubisco. Proc Natl Acad Sci USA 102:17225-17230