MOLECULAR CLONING AND EXPRESSION OF THE PROLIFERATING CELL NUCLEAR ANTIGEN GENE FROM THE COCCOLITHOPHORID PLEUROCHRYSIS CARTERAE (HAPTOPHYCEAE)1

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Abstract
The gene encoding proliferating cell nuclear antigen (PCNA) was isolated from the marine coccolithophorid microalga Pleurochrysis carterae (Braarud et Fagerland) Christensen (Haptophyceae). Two mRNAs (Pcpcna1 and Pcpcna2) were identified and contained an identical coding region for 222 amino acid residues and an untranslated sequence of 302 base pair (Ut1) and 246 base pair (Ut2), respectively. Comparison between PCR‐derived genomic DNA fragments and cDNA sequences revealed five introns. The coding region of Pcpcna is similar to counterparts in other organisms and contains highly conserved functional domains. Phylogenetic analyses indicated clustering of Pcpcna with pcna in its haptophyte relative Isochrysis galbana Parke. A recombinant fusion protein of Pcpcna, overexpressed in Escherichia coli, was recognized by the PC10 antibody against rat PCNA. Using RT‐PCR and Western blotting, Pcpcna was found to be highly transcribed and translated during the exponential growth phase relative to the stationary growth phase, with a positive correlation between gene expression and growth rate. It can be concluded that the pcna is conserved in this coccolithophorid phytoplankton and that its expression is growth stage related.

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