CC-4877 rbcL-C247S mt+
$30.00
From Robert J. Spreitzer, University of Nebraska, August 2014
Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆-MX3312 mt+ (CC-4696) (Satagopan and Spreitzer 2004), a C247S substitution was created in the Rubisco large subunit. This mutant was created to investigate phylogenetic differences in the Rubisco large subunit that may be responsible for pyrenoid formation in algae (Nozaki et al. 2002). See also rbcL-R32K/A222S/C247S. The C247S substitution does not affect Rubisco catalysis or pyrenoid formation (Lim and Meyer, unpublished). Cys-247 forms a disulfide bond with Cys-247 in a neighboring large subunit, which may be the only disulfide bond in the Rubisco holoenzyme (Knight et al. 1990). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.
Knight S, Andersson I, Branden CI (1990) Crystallographic analysis of ribulose 1,5-bisphosphate carboxylase from spinach at 2.4 A resolution. Subunit interactions and active site. J Mol Biol 215:113-160
Nozaki H, Onishi K, Morita E (2002) Differences in pyrenoid morphology are correlated with differences in the rbcL genes of members of the Chloromonas lineage (volvocales, chlorophyceae). J Mol Evol 55:414-430
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