CC-4730 rbcS1-R59E pf2 mt+
$30.00
From Robert J. Spreitzer, University of Nebraska, July 2014
Phenotype: requires acetate at 35 °C, temperature-conditional
Using a plasmid that would create an R59E substitution (CGC-GAG) in the betaA-betaB loop of the Rubisco small subunit (Spreitzer et al. 2001; Spreitzer 2003), Kelsey Swartz in Spreitzer’s group transformed rbcS∆-CAL005.01.13 pf2 mt+ (CC-4692) (Dent et al. 2005). In contrast to the R59E mutant described previously (Spreitzer et al. 2001), this strain has a cell wall and can be used in genetic crosses. This mutant can grow on minimal medium at 25 °C, but dies on minimal medium at 35 °C. The strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function or stability.
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
Spreitzer RJ (2003) Role of the small subunit in ribulose-1,5-bisphosphate carboxylase/oxygenase. Arch Biochem Biophys 414:141-149
Spreitzer RJ, Esquivel MG, Du YC, McLaughlin PD (2001) Alanine-scanning mutagenesis of the small-subunit betaA-betaB loop of chloroplast ribulose-1,5-bisphosphate carboxylase/oxygenase: Substitution at Arg-71 affects thermal stability and CO2/O2 specificity. Biochemistry 40:5615-5621