Protistology 5 (2/3) 131-141 (2007/8)
Macroarray-based gene expression analysis in Tetrahymena pyriformis
Bernhard F. Benkel, Scott Richmond, Jon Davoren, Michael Ivan, Ronald M. Teather and Robert J. Forster
Agriculture and Agri-Food Canada, Lethbridge Research Centre, Lethbridge, Alberta, Canada
Summary
A cDNA library was constructed for the aquatic ciliate Tetrahymena pyriformis, and one hundred
and fifty-three clones we reused for gene expression analysis in a filter array hybridization assay. Expression
levels for individual genes ranged from easily detectable to below the detection threshold of the assay. Ten cDNAs
showed relatively intense hybridization signals, and two of these were chosen for further analysis. One of these
genes, Tp177, was characterized in detail, i.e. its transcribed and putative promoter regions were completely
defined and analyses carried out to identify potential gene regulatory elements. Partial nucleotide sequences were
derived from fifty-seven cDNA clones and compared to the GenBank sequence database.Twenty-six of these clones showed
significant similarity to genes in GenBank (eleven of these represented ribosomal proteins), while the remainder,
including clone Tp177, represented novel sequences.
Key words: EST, cDNA array, transcript abundance, gene cloning, long range-inverse PCR,
DNA sequencing, protozoa
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