Identification and spatial distribution of the mRNA encoding the gp49 component of the gilthead sea bream, Sparus aurata, egg envelope

A cDNA encoding the precursor of one of the major components of gilthead sea bream, Sparus aurata, egg envelope has been cloned by reverse transcriptase polymerase chain reaction (RT-PCR) techniques. The clone was isolated starting from total RNA extracted from the liver of spawning female fish and estradiol-17 beta-treated male fish. Sequence analysis revealed that the cDNA encoded a protein of 405 aa corresponding to 49-kDa component (termed gp49), a glycoprotein belonging to the N-linked type. The gp49 protein is homologous to the ZI-3 of medaka Oryzias latipes, the mammalian ZPC and ZPC homologues of Xenopus laevis (xlZPC) and carp Cyprinus carpio (ccZPC). In addition, the open reading frame also encodes an additional aa sequence, the signal peptide, located in the N terminal region of the protein. RT-PCR and in situ expression analyses evidenced an organ-restricted pattern: the mRNA was detected only in liver of spawning female and estradiol-17 beta-treated male fish but not in other tissues.