CC-4933 rbcL-LSAT16 mt+
$30.00
From Robert J. Spreitzer, University of Nebraska, October 2014
Fuqiao Xu in Spreitzer’s group created a codon-optimized Arabidopsis rbcL gene that encodes a large subunit with 12 Chlamydomonas amino-acid substitutions (S2V, S10G, V11A, E19D, K21R, N442G, E443D, I444V, E447S, T466K, N468E, P470D) and that lacks four C-terminal residues (Asp-476, Gly-477, Gln-478, Glu-479). When the engineered gene was transformed into rbcL∆-MX3312 mt+ (CC-4696), photosynthesis-competent transformants were recovered on minimal medium with 5% CO2 in air. Thus, this mutant expresses a functional Rubisco comprised of Chlamydomonas small subunits and engineered Arabidopsis large subunits. The mutant grows less than wild type on minimal medium, but the same as wild type on minimal medium with 5% CO2 in air. Western analysis indicated that the mutant strain has a normal level of Rubisco holoenzyme (Xu and Spreitzer, unpublished). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.