Molecular cloning of a cDNA encoding vitellogenesis-inhibiting hormone in the whiteleg shrimp Litopenaeus vannamei and preparation of its recombinant peptide using an E. coli expression system

• Published in: Fisheries science Vol. 79; no. 3; pp. 357 - 365
• Main Authors: Tsutsui, Naoaki, Ohira, Tsuyoshi, Okutsu, Tomoyuki, Shinji, Junpei, Bae, Sun-Hye, Kang, Bong Jung, Wilder, Marcy N.
• Format: Journal Article
• Language: English
• Published: Japan Springer Japan 01.05.2013; Springer Nature B.V
• Subjects: Amino acids; Bioassays; Biological activity; Biomedical and Life Sciences; Cloning; Crustaceans; Deoxyribonucleic acid; DNA; E coli; Escherichia coli; Fish & Wildlife Biology & Management; Fisheries; Fishing; Food Science; Freshwater & Marine Ecology; Gene expression; Life Sciences; Litopenaeus vannamei; Original Article; Ovaries; Peptides; Science; Seafood; Shellfish; Sinuses; Studies; Japan; Whiteleg shrimp; Crustacean hyperglycemic hormone family; Vitellogenesis-inhibiting hormone; Vitellogenin gene
• ISSN: 0919-9268; 1444-2906
• DOI: 10.1007/s12562-013-0603-z

The cDNA encoding a precursor of Liv-SGP-G, the most abundant crustacean hyperglycemic hormone-family peptide in the sinus gland of the whiteleg shrimp Litopenaeus vannamei , was cloned by RT-PCR coupled with 5′- and 3′-RACE. The determined cDNA sequence consisted of 621 nucleotides comprising a 69-bp 5′-untranslated region, a 357-bp open reading frame, and a 195-bp 3′-untranslated region. The deduced sequence of 72 amino acid residues corresponding to mature Liv-SGP-G was completely identical to that of natural Liv-SGP-G, determined in our previous study. The recombinant Liv-SGP-G was thereafter heterologously expressed in Escherichia coli , and its vitellogenesis-inhibiting activity in ex vivo cultured ovarian fragments was examined. The recombinant peptides inhibited vitellogenin gene expression with almost the same efficacy as that of natural Liv-SGP-G purified from sinus glands.