Subategory:

Mutants

Description:

Using plasmid p699, rbcL direct mutagenesis, chloroplast transformation of wild-type 2137 mt+ (cloned), selection for spectinomycin resistance in the dark, and screening for a homoplasmic acetate-requiring phenotype to create a K175R substitution (AAA-CGT) in the Rubisco large subunit were performed. The K175R substitution causes a 50% decrease in Rubisco holoenzyme and a 99% decrease in Rubisco carboxylase activity. The x-ray crystal structure of this K175R Rubisco enzyme has been solved (Andersson and Spreitzer, unpublished). This mutant was created to investigate the role of active-site Lys-175 in catalysis (Harpel et al. 2002). It has been maintained with acetate medium in darkness since its creation.

Species:

Chlamydomonas

Locus:

rbcL

Chromosome:

Chloroplast

Phenotype:

Requires acetate; sensitive to light