CC-4813 rbcL-L326I/M349L mt+
$30.00
From Robert J. Spreitzer, University of Nebraska, July 2014
Directed mutagenesis and chloroplast transformation of rbcL∆-25B1 mt+ (CC-4700) were used to create L326I (CTT-ATT) and M349L (ATG-CTG) substitutions in the Rubisco large subunit, which complemented for structural stability but decreased CO2/O2 specificity (Zhu and Spreitzer 1996). The L326I/M349L double mutant was created to investigate phylogenetic differences near large-subunit residue Val-331 (see rbcL-V331A) (Chen and Spreitzer 1989; Chen et al. 1991). After being maintained with acetate medium in darkness for several years, the original L326I/M349L double-mutant culture was found to contain only two types of mutant cells, both of which had additional mutations in the rbcL gene (Satagopan et al. 2000). L326I/V341I/M349L/A378T mutant cells had an acetate-requiring phenotype, and H310N/L326I/M349L cells had a wild-type phenotype. It was proposed that these additional substitutions may have been selected because they improve Rubisco holoenzyme stability (Satagopan et al. 2000). See also CC-4713 and CC-4714. Yvonne Arendsen in Spreitzer’s group created the rbcL-L326I/M349L mt+ mutant again in 2000, and that is the strain listed here. It has been maintained with acetate medium in darkness since it was created, but it has not been investigated for the presence of suppressor mutations.
Chen Z, Spreitzer RJ (1989) Chloroplast intragenic suppression enhances the low CO2/O2 specificity of mutant ribulosebisphosphate carboxylase/oxygenase. J Biol Chem 264:3051-3053
Chen Z, Yu W, Lee JH, Diao R, Spreitzer RJ (1991) Complementing amino-acid substitutions within loop 6 of the alpha/beta-barrel active site influence the CO2/O2 specificity of chloroplast ribulose-1,5-bisphosphate carboxylase/oxygenase. Biochemistry 30:8846-8850
Satagopan S, Arendsen YJ, Spreitzer RJ (2000) Selection for improved Rubisco stability in the absence of photosynthesis. Plant Biology 2000:809
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