CC-4941 rbcL-C256A/C369A 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), two substitutions (C256A and C369A) were created together in the Rubisco large subunit. Because Cys-256 and Cys-369 are methylated (Taylor et al. 2001), this mutant was created to investigate the role of the modified residues in Rubisco structure or function. The C256A/C369A substitutions cause decreases in Rubisco CO2/O2 specificity and carboxylation catalytic efficiency (Spreitzer et al., unpublished).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
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