CC-4716 rbcL-C449S/C459S/K474C mt+
$30.00
From Robert J. Spreitzer, University of Nebraska, July 2014
Girish Rasineni in Spreitzer’s group performed directed mutagenesis of plasmid pLS-C449S/C459S (Marin-Navarro and Moreno 2006), obtained from Joaquin Moreno in 2011, to produce an additional K474C substitution (AAA-TGT). The triple-mutant plasmid was then transformed into rbcL∆-MX3312 mt+ (CC-4696) by standard procedures (Zhu and Spreitzer 1996), and photosynthesis-competent colonies were selected on minimal medium. After many rounds of cloning for homoplasmicity, this strain was retained for future study. It is maintained with acetate medium in darkness to prevent selection of other mutations.
Marin-Navarro J, Moreno J (2006) Cysteines 449 and 459 modulate the reduction–oxidation conformational changes of ribulose 1·5-bisphosphate carboxylase/oxygenase and the translocation of the enzyme to membranes during stress. Plant Cell Environ 29:898-908
Zhu G, Spreitzer RJ (1996) Directed mutagenesis of chloroplast ribulose-1,5-bisphosphate carboxylase/oxygenase: Loop-6 substitutions complement for structural stability but decrease catalytic efficiency. J Biol Chem 271:18494-18498