Molecular cloning of a molt-inhibiting hormone cDNA from the kuruma prawn Penaeus japonicus

Zoological Science
Tsuyoshi OhiraK Aida

Abstract

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 positive 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.

Citations

Apr 4, 2000·Insect Biochemistry and Molecular Biology·H EndoT Watanabe
Feb 23, 2012·International Journal of Biological Sciences·Philip BradyWayne Knibb
Nov 22, 2001·Biochemical and Biophysical Research Communications·W J Yang, K R Rao
Jun 9, 2007·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Hideaki AsazumaHiromichi Nagasawa
May 14, 2005·Annals of the New York Academy of Sciences·Hideaki AsazumaHiromichi Nagasawa
Sep 10, 1998·Biochemical and Biophysical Research Communications·K J LeeR D Roer
Nov 13, 2008·Comparative Biochemistry and Physiology. Part A, Molecular & Integrative Physiology·Teruaki NakatsujiR Douglas Watson
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Methods Mentioned

BETA
PCR
electrophoresis

Software Mentioned

Clustul V

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