Abstract
Pleurochrysis haptonemofera is a unicellular marine coccolithophorid that has calcified scales, coccoliths, on the cell surface. Some coccolithophorids including P. haptonemofera have a coccolith-bearing stage and a naked stage in their life cycles. To characterize genes involved in the coccolithogenesis, we generated a total of 9550 expressed sequence tags (EST) from a normalized cDNA library that was prepared using both coccolith-bearing cells (C-cells) and naked cells (N-cells), constructed a cDNA macroarray using the EST clones, and then analyzed the gene expression specificity in C-cells and N-cells. When cDNA clones whose expression ratio exceeded 3-fold were selected, as many as 180 clones were identified as C-cell-specific ones, while only 12 were found to be N-cell-specific ones. These clones were sequenced, assembled, and homology-searched against a public nonredundant protein database. As a result, they were grouped into 54 C-cell-specific and 6 N-cell-specific genes, and 59% and 50% of these genes exhibited significant similarity to those of other known proteins, respectively. To assess mRNA expression further, Northern hybridization was performed for 12 of the C-cell-specific genes and one of the N-cell-specific ones. These clones, together with the new cDNA macroarray, will provide a powerful tool for the future genome-wide functional analysis of uncharacterized genes related to the regulation of the calcification and life cycle of coccolithophorids.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Animals
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Base Sequence
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Blotting, Northern / veterinary
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DNA, Protozoan / analysis
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DNA, Protozoan / biosynthesis
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DNA, Protozoan / chemistry
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Eukaryota / genetics*
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Eukaryota / physiology
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Gene Expression Profiling / veterinary*
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Genes, Protozoan / genetics*
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Genes, Protozoan / physiology
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Molecular Sequence Data
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Oligonucleotide Array Sequence Analysis / veterinary*
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Sequence Alignment / veterinary
Associated data
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GENBANK/AB258464
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GENBANK/AB258465
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GENBANK/AB258466
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GENBANK/AB258467
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