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1 November 2009 Localization and Copy Number of the Protein-Coding Genes Actin, α-Tubulin, and HSP90 in the Nucleus of a Primitive Dinoflagellate, Oxyrrhis marina
Junji Sano, Koichi H. Kato
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Abstract

Chromosomes of the dinoflagellate Oxyrrhis marina are composed of thin parallel filaments running longitudinally and lack the arched structure common to the dinochromosome. The physicochemical and molecular organization of these chromosomes, including the localization and arrangement of genes, is still unknown. We investigated the locations of three protein-coding genes, actin (AF482402), α-tubulin (AF482403), and HSP90 (AY391258), on these chromosomes using fluorescence in-situ hybridization (FISH). Primers for these three genes were designated according to known partial sequences. PCR products amplified from total DNA were labeled with digoxigenin (DIG) by random priming and used as probes. After in-situ hybridization, DIG signals were amplified and visualized with anti-DIG-FITC. The number of signals was 3±1.3 (n=90) for actin, 4.1±1.4 (n=70) for α-tubulin, and 5.5±1.7 (n=80) for HSP90. This study is the first to locate protein-coding genes in the nucleus of a dinoflagellate, although the chromosomes were greatly damaged during the FISH process. The copy number of each gene per cell was estimated using real time PCR. Resulting copy numbers of actin, α-tubulin and HSP90 were, 33.7, 10.4 and 5.4, respectively.

© 2009 Zoological Society of Japan
Junji Sano and Koichi H. Kato "Localization and Copy Number of the Protein-Coding Genes Actin, α-Tubulin, and HSP90 in the Nucleus of a Primitive Dinoflagellate, Oxyrrhis marina," Zoological Science 26(11), 745-753, (1 November 2009). https://doi.org/10.2108/zsj.26.745
Received: 11 February 2009; Accepted: 20 July 2009; Published: 1 November 2009
KEYWORDS
actin gene
dinoflagellate
fluorescence in-situ hybridization (FISH)
HSP90 gene
Oxyrrhis marina
α-tubulin gene
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