Isolation of Cycloheximide-resistant Mutants of Cyanidioschyzon merolae

Cyanidioschyzon merolae is a unicellular red alga that lives in acidic hot springs. The genome sequence of C. merolae has been completely read, but a lack of transformation systems still limits its application in genetics. To choose an appropriate drug for use in selectable media, we examined the effects of seven antibiotics on the growth of C. merolae. Only cycloheximide, an inhibitor of protein synthesis, effectively inhibited the growth. We noticed that there was a population that could survive in the presence of cycloheximide and succeeded in isolating six cycloheximide-resistant clones. All these clones have the same single mutation in the ribosomal protein L29 gene that encodes a ribosomal protein.

[1]  Fumiko Ohta,et al.  Genome sequence of the ultrasmall unicellular red alga Cyanidioschyzon merolae 10D , 2004, Nature.

[2]  T. Kuroiwa,et al.  A Plant-Specific Dynamin-Related Protein Forms a Ring at the Chloroplast Division Site Article, publication date, and citation information can be found at www.plantcell.org/cgi/doi/10.1105/tpc.009373. , 2003, The Plant Cell Online.

[3]  T. Kuroiwa.  The primitive red algae Cyanidium caldarium and Cyanidioschyzon merolae as model system for investigating the dividing apparatus of mitochondria and plastids , 1998 .

[4]  T. Kuroiwa,et al.  Comparison of Ultrastructures between the Ultra-small Eukaryote Cyanidioschyzon merolae and Cyanidium caldarium , 1994 .

[5]  T. Kuroiwa,et al.  Behavior of mitochondria, chloroplasts and their nuclei during the mitotic cycle in the ultramicroalga Cyanidioschyzon merolae. , 1994, European journal of cell biology.

[6]  M. Yao,et al.  Transformation of Tetrahymena to cycloheximide resistance with a ribosomal protein gene through sequence replacement. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[7]  A. Böck,et al.  Amino acid replacements in ribosomal protein YL24 of Saccharomyces cerevisiae causing resistance to cycloheximide , 1981, FEBS letters.

[8]  M. B. Allen.  Studies with cyanidium caldarium, an anomalously pigmented chlorophyte , 2004, Archiv für Mikrobiologie.

[9]  K. Toda.  Molecular cytological studies of nuclear shapes and chromosomes in an ultra-small primitive red alga, Cyanidioschyzon merolae , 1996 .