Subategory:

Mutants

Description:

A codon-optimized Arabidopsis rbcL gene was created 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 rbcLΔ-MX3312 mt+ (MUT-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. This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.

Species:

Chlamydomonas

Locus:

rbcL

Chromosome:

Chloroplast