CC-4864 rbcS1-L66F mt
$30.00
From Robert J. Spreitzer, University of Nebraska, August 2014
Phenotype: requires acetate at 35 °C, temperature-conditional
Using standard methods of directed mutagenesis and nuclear-gene transformation of rbcS∆-T60-3 mt- (CC-4690) (Genkov and Spreitzer 2009), Todor Genkov in Spreitzer’s group created an L66F substitution (CTG-TTC) in the Rubisco small subunit. This mutant was created to see whether an L66F substitution in the nuclear-encoded small subunit would mimic the biochemical defects that arise from an L290F substitution in the chloroplast-encoded large subunit (Chen et al. 1988). Like mutant rbcL-L290F (68-4PP), the rbcS1-L66F mutant has a temperature-conditional phenotype. It grows on minimal medium at 25 °C, but dies on minimal medium at 35 °C (Genkov and Spreitzer, unpublished). L66F Rubisco also has decreases in CO2/O2 specificity and holoenzyme stability in vivo and in vitro (Genkov and Spreitzer, unpublished). See also rbcS1-L66F pf2 mt+ (CC-4737). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.
Chen Z, Chastain CJ, Al-Abed SR, Chollet R, Spreitzer RJ (1988) Reduced CO2/O2 specificity of ribulose-1,5-bisphosphate carboxylase/oxygenase in a temperature-sensitive chloroplast mutant of Chlamydomonas reinhardtii. Proc Natl Acad Sci USA 85:4696-4699
Genkov T, Spreitzer RJ (2009) Highly conserved small subunit residues influence Rubisco large subunit catalysis. J Biol Chem 284:30105-30112