Molecular cloning and characterization of three distinct choriogenins in masu salmon, Oncorhynchus masou

• Published in: Molecular reproduction and development Vol. 75; no. 7; pp. 1217 - 1228
• Main Authors: Fujita, Toshiaki, Fukada, Haruhisa, Shimizu, Munetaka, Hiramatsu, Naoshi, Hara, Akihiko
• Format: Journal Article
• Language: English
• Published: Hoboken Wiley Subscription Services, Inc., A Wiley Company 01.07.2008; Wiley-Liss
• Subjects: Amino Acid Sequence; Animals; Biological and medical sciences; Biotechnology; choriogenesis; Cloning, Molecular; DNA Primers; DNA, Complementary - genetics; Egg Proteins - genetics; Female; Fundamental and applied biological sciences. Psychology; Genetic engineering; Genetic technics; heterodimerization; Methods. Procedures. Technologies; Molecular cloning; Molecular Sequence Data; Nucleic Acid Hybridization; Oncorhynchus - classification; Oncorhynchus - genetics; Oncorhynchus masou; Phylogeny; Protein Precursors - genetics; recombinant protein; Recombinant Proteins - analysis; Sequence Alignment; Sequence Homology, Amino Acid; teleost; Oncorhynchus masou; Salmon; Chorion; teleost; Germinal cell; Heterodimer; Zona pellucida; Characterization; Vertebrata; Pisces; heterodimerization; Recombinant protein; choriogenesis; Molecular cloning; Oocyte
• ISSN: 1040-452X; 1098-2795
• DOI: 10.1002/mrd.20857

Three cDNAs, each encoding a different choriogenin (Chg), were isolated from a female masu salmon (Oncorhynchus masou) liver cDNA library. Two of the cDNA clones, Chg Hα and Chg Hβ, showed a close relationship and contained the typical domains of zona pellucida (ZP) B genes in fish, namely proline and glutamine rich repeats, a trefoil factor family domain, and a ZP domain. Specific antibodies against recombinant Chg H products (rmHα and rmHβ) were generated to elucidate the relationship between the Chg H cDNAs and two types of serum Chg H protein, which were previously purified and characterized, and designated as very‐high‐molecular‐weight vitelline envelope‐related protein (vhVERP) and Chg H of masu salmon. The immunobiochemical analyses revealed that the Chg Hα and Chg Hβ clones encoded vhVERP and Chg H proteins, respectively. The third cDNA clone (Chg L) appeared to be a ZPC gene and, by mapping the N‐terminal sequence of purified Chg L, was shown to encode serum Chg L protein. Various types of heteromultimer of the three Chgs were identified immunologically as high molecular weight chorion components, indicating the involvement of complex heterodimerization of multiple Chgs in the construction of chorion architecture in masu salmon. Mol. Reprod. Dev. 75: 1217–1228, 2008. © 2008 Wiley‐Liss, Inc.