CC-5026 rbcL-AG86 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 six substitutions (V30E, V31T, R32K, M42V, C53A, and D86H,) together in the Rubisco large subunit. This mutant (also named AG86) 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 The mutant enzyme has a decrease in CO2/O2 specificity (Lim and Spreitzer, unpublished). The mutant strain has a small decrease in photosynthetic growth at 35 °C. 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
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