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Updated: Aug 2, 2026

High-Throughput Robotically Assisted Isolation of Temperature-sensitive Lethal Mutants in Chlamydomonas reinhardtii
Published on: December 5, 2016
Mutations in nine chloroplast loci of Chlamydomonas affecting different photosynthetic functions
Abstract:
Chloroplast components known to be coded by chloroplast DNA include chloroplast rRNAs, tRNAs, and the large subunit of ribulose-bisphosphate carboxylase. Because these components comprise less than 3% of the estimated coding capacity of the chloroplast genome, most chloroplast gene functions have yet to be identified. One approach to this problem is the isolation and characterization of mutations in the chloroplast genome affecting specific photosynthetic functions. Recently we have found that such mutations can be preferentially recovered by using arsenate selection on cells previously grown in 5-fluorodeoxyuridine. Sixteen mutants thus isolated have been localized into nine chloroplast loci, based on their ability to recombine and produce photosynthetically competent progeny. Mutants at two loci show the characteristic syndrome of photosynthetic defects that results from a deficiency in chloroplast protein synthesis. These have been found to lack chloroplast ribosome monomers. Mutants at three loci are missing chlorophyll-protein complex I in their thylakoid membranes. Mutants at three other loci are deficient in membrane polypeptides known to be associated with the chloroplast coupling factor.
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