CC-4939 rbcL-C256A mt+
$30.00
From Robert J. Spreitzer, University of Nebraska, November 2014
Phenotype: requires acetate at 35 °C, temperature-conditional
Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆-MX3312 mt+ (CC-4696) (Satagopan and Spreitzer 2004), a C256A substitution (TGT-GCT) was created in the Rubisco large subunit. Because Cys-256 is methylated (Taylor et al. 2001), this mutant was created to investigate the role of the modified residue in Rubisco structure or function. Large-subunit Cys-256 has significant interactions with the Rubisco small subunit (Spreitzer et al. 2005). The C256A substitution causes decreases in Rubisco CO2/O2 specificity and carboxylation catalytic efficiency (Spreitzer et al., unpublished). The mutant strain grows slowly on minimal medium at 25 °C, but dies on minimal medium at 35 °C. This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.
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
Taylor TC, Backlund A, Bjorhall K, Spreitzer RJ, Andersson I (2001) First crystal structure of Rubisco from a green alga, Chlamydomonas reinhardtii. J Biol Chem 276:48159-48164