Fig. 2
Cloned cDNA encoding mZPC5 in the ovaries and livers of spawning medakas. A: the genomic structure of mzpc5 and the primer position used to identify mzpc5 expression and clone full-length cDNA. The diagrammed pattern of the genomic structure is based on information from the Ensemble Project database [36]. The blue and red arrows indicate the positions of forward (F1, F2, F3) and reverse (R1, R2, and R3) primers, respectively, that were used for RT-PCR. B: the result of the RT-PCR analysis. Ba: RNA obtained from the spawning female ovary. Bb: RNA obtained from the spawning female liver. Bc: RNA obtained from male liver. C: the nucleotide and predicted amino acid sequences of newly cloned mzpc5 cDNA in this study. A one-letter symbol represents each amino acid as defined by the International Union of Pure and Applied Chemistry (IUPAC). The red arrowhead points to the signal sequence cleavage site analyzed using PSORT [37]. Underlined amino acid sequences with bold red letters represent the antigen peptide sequence. The blue and red arrows indicate the locations of primers used for RT-PCR analysis. Blue-circled letters indicate the predicted amino acids connecting to O-linked oligosaccharides [38]. Letters boxed in blue represent the predicted peptide sequence at the N-glycosylation site [39]. Bold blue amino acids (312G and 343 K) differ from the amino acids in ZPC5 (accession # AAN31192). The amino acid sequences of the peptides shown in bold black letters were obtained from the immunoreactive proteins (~74 kDa) in Figs. 4b and 5b following MS/MS analysis