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Dinoflagellates are an important group of unicellular eukaryotes widespread in aquatic ecosystems. Many dinoflagellates are mixotrophic, toxic or potentially toxic, highly competitive and invasive, while molecular mechanisms that underpin their success in natural communities remain enigmatic. Due to peculiar features of dinoflagellate genome, little is known about the structure and expression of genes in these organisms. We analyzed the transcriptome databases of the dinoflagellate Prorocentrum minimum in order to identify the sequences of urea transporter (dur3) and nitrate transporter (nrt2.1) genes. Taking into account prospective exon-intron organization of dinoflagellate genome we suggested two variants of choosing the primer positions. We designed six primer pairs for amplification of the urea transporter gene fragments and three – for amplification of the nitrate transporter gene fragments. As a result of PCR, fragments of target genes were obtained. Alignment of amplicons with database transcriptome sequences showed that those sequences were identical. Primers developed in this study can be further used for examination of P. minimum gene expression by RT-qPCR. This approach would provide a better understanding of the influence of various nitrogen sources on physiological characteristics of these protists responsible for their effective adaptations to fluctuating environment.

Molecular tools for invasion biology: a new approach for amplification of dinoflagellate nitrogen transport genes with unknown exon-intron structure