Determination of amino acid sequence and site of mRNA expression of four vitellins in the giant freshwater prawn, Macrobrachium rosenbergii

Wei Jun Yang, Tsuyoshi Ohira, Naoaki Tsutsui, Thanumalayaperumal Subramoniam, Do Thi Thanh Huong, Katsumi Aida, Marcy N. Wilder

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Four major yolk proteins, designated as vitellins (Vns) Macr-VnA, B, C, and D, were extracted from mature ovaries of Macrobrachium rosenbergii. These were purified to homogeneity by reversed-phase high performance liquid chromatography (HPLC) employing a unique separation system based on the hydrophobic properties of the Vn molecule. Using standard techniques of protein sequencing, more than 33 N-terminal and 57 internal amino acid residues were determined for each of the four Vns. The cDNA fragments encoding the four Vns were amplified by PCR using degenerate oligonucleotide primers derived from the N-terminal and internal amino acid sequences. These cDNA fragments were cloned, sequenced, and used as probes to examine the transcription of mRNAs encoding the four Vns. Significant accumulations of these mRNAs were observed in female hepatopancreas only, while mRNA expression was not detected in male hepatopancreas or any other female tissue including ovary, subepidermal adipose tissue, gill, and muscle. This is the first occasion in Crustacea in which multiple Vns were demonstrated to be synthesized simultaneously in a single tissue. (C) 2000 Wiley-Liss, Inc.

Original languageEnglish
Pages (from-to)413-422
Number of pages10
JournalJournal of Experimental Zoology
Issue number6
Publication statusPublished - Nov 1 2000


ASJC Scopus subject areas

  • Animal Science and Zoology

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