CC-4737 rbcS1-L66F pf2 mt+
$30.00
From Robert J. Spreitzer, University of Nebraska, July 2014
Phenotype: requires acetate at 35 °C, temperature-conditional
Directed mutagenesis and nuclear-gene transformation of rbcS∆-CAL005.01.13 pf2 mt+ (CC-4692) (Dent et al. 2005; Genkov et al. 2006) were used to create an L66F substitution (CTG-TTC) in the betaA-betaB loop of the Rubisco small subunit (Genkov and Spreitzer, unpublished), which mimics the biochemical defects of chloroplast Rubisco mutant rbcL-L290F (Chen et al. 1988). This mutant can grow on minimal medium at 25 °C, but dies on minimal medium at 35 °C. It has a cell wall and can be used in genetic crosses. The strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function or stability.
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
Dent RM, Haglund CM, Chin BL, Kobayashi MC, Niyogi KK (2005) Functional genomics of eukaryotic photosynthesis using insertional mutagenesis of Chlamydomonas reinhardtii. Plant Physiol 137:545-556
Genkov T, Du YC, Spreitzer RJ (2006) Small-subunit cysteine-65 substitutions can suppress or induce alterations in the large-subunit catalytic efficiency and holoenzyme thermal stability of ribulose-1,5-bisphosphate carboxylase/oxygenase. Arch Biochem Biophys 451:167-174