CC-4934 rbcL-LSAT16/rbcS1-SSAT pf2 mt+
$30.00
From Robert J. Spreitzer, University of Nebraska, October 2014
Phenotype: requires elevated CO2 for photosynthetic growth
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∆/rbcS1-SSAT pf2 mt+ (which lacks Chlamydomonas rbcL and rbcS, but expresses Arabidopsis rbcS), photosynthesis-competent transformants were recovered on minimal medium with 5% CO2 in air. Thus, this mutant expresses a functional Rubisco comprised of Arabidopsis small subunits and engineered Arabidopsis large subunits. On minimal medium with 5% CO2 in air, the mutant grows less than rbcL-LSATNC mt+ and rbcL-LSATNC-T391F mt+, but more than rbcL-LSATNC-T391F/rbcS1-SSAT pf2 mt+. Western analysis indicated that the mutant strain has a normal level of Rubisco holoenzyme (Xu and Spreitzer, unpublished).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.
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