CC-5035 RbcL-AG305-474 Mt+ - Chlamydomonas Resource Center

CC-5035 rbcL-AG305-474 mt+

$30.00

From Robert J. Spreitzer, University of Nebraska, November 2014

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆-MX3312 mt+ (CC-4696) (Satagopan and Spreitzer 2004), Boon Hoe Lim in Spreitzer’s group created eight substitutions (M42V, C53A, R305K, V341I, D470E, T471A, I472M, and K474T) together in the Rubisco large subunit. This mutant (also named AG305-474) 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). This strain 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

Satagopan S, Spreitzer RJ (2004) Substitutions at the Asp-473 latch residue of Chlamydomonas ribulosebisphosphate carboxylase/oxygenase cause decreases in carboxylation efficiency and CO2/O2 specificity. J Biol Chem 279:14240-14244

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

• Locus:
• rbcL
• Chromosome:
• chloroplast

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