CC-4945 rbcL-P104A/P151A/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), Girish Rasineni in Spreitzer’s group created four substitutions together (P104A, P151A, C256A, and C369A) in the Rubisco large subunit. Because Pro-104 and Pro-151 are hydroxylated, and Cys-256 and Cys-369 are methylated (Taylor et al. 2001), the mutant was created to investigate the role of the modified residues in Rubisco structure or function. The four substitutions cause decreases in Rubisco CO2/O2 specificity and carboxylation catalytic efficiency (Rasineni and Spreitzer, unpublished).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