Molecular Cloning of a Molt-Inhibiting Hormone cDNA from the Kuruma Prawn Penaeus japonicus

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The crustacean molt-inhibiting hormone (MIH) is released from the X-organ sinus gland complex and suppresses ecdysteroid synthesis by the Y-organ. In the present study, we have isolated a cDNA which encodes a MIH (Pej-SGP-IV) of the kuruma prawn Penaeus japonicus in order to study its expression and characterize the structure of its precursor. A cDNa fragment was isolated using RT-PCR with two degenerate oligonucleotide primers that were designed based on the peptide sequence of Pej-SGP-IV, and this fragment was used as a probe to screen an eyestalk cDNa library. In a posotive cDNA clone (814 base pairs (bp)), an open reading frame of 315 bp was found ; the conceptually translated protein consists of a putative signal peptide (28 residues) and Pej-SGP-IV (77 residues). In Northern blot analysis using a cDNA probe, specific hybridization to a transcript of 0.95 kb was seen in RNA extracted from the eyestalk but not from hepatopancreas, abdominal muscle, brain, thoracic ganglia or abdominal ganglia. The level of the Pej-SGP-IV mRNA in the eyestalk did not change significantly during the molt cycle.
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