CC-4743 rbcS1-ABEFGP mt-
$30.00
From Robert J. Spreitzer, University of Nebraska, July 2014
Phenotype: requires acetate at 35 °C, temperature-conditional
Using standard methods of genetic engineering and nuclear-gene transformation of rbcS∆-T60-3 mt- (CC-4690) (Genkov et al. 2010), the betaA-betaB loop of the Rubisco small subunit was replaced with the small-subunit betaA-betaB loop from Galdieria partita, and residues 128 through 140 were replaced with the small-subunit betaE-betaF loop (residues 105 through 138) from Galdieria. These changes cause a decrease in Rubisco CO2/O2 specificity and holoenzyme stability (Genkov and Spreitzer, unpublished). The mutant can grow on minimal medium at 25 °C, but dies on minimal medium at 35 °C. This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function or stability.
Genkov T, Meyer M, Griffiths H, Spreitzer RJ (2010) Functional hybrid Rubisco enzymes with plant small subunits and algal large subunits: Engineered rbcS cDNA for expression in Chlamydomonas. J Biol Chem 285:19833-19841