Expression of a recombinant molt-inhibiting hormone of the kuruma prawn Penaeus japonicus in Escherichia coli.
The crustacean molt-inhibiting hormone (MIH) suppresses ecdysteroid synthesis by the Y-organ. The MIH of the kuruma prawn Penaeus japonicus has recently been isolated and its cDNA cloned. In this study, we expressed the MIH in Escherichia coli to obtain a large quantity of this hormone with biological activity. The MIH cDNA was processed and ligated into an expression plasmid. E. coli was transformed with this plasmid, and then the recombinant MIH (r-MIH) was expressed. The r-MIH was put through the refolding reaction and was purified by reverse-phase HPLC. N-terminal amino acid sequence and time-of-flight mass spectral analyses supported the idea that the r-MIH had the entire sequence. By in vitro bioassay using the Y-organ of the crayfish, the r-MIH was found to be comparable to natural MIH in inhibiting ecdysteroid synthesis.
Ohira
T
T
Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, University of Tokyo, Japan.
Nishimura
T
T
Sonobe
H
H
Okuno
A
A
Watanabe
T
T
Nagasawa
H
H
Kawazoe
I
I
Aida
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D007447:Invertebrate Hormones / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D033561:Penaeidae / Q000235:genetics* / Q000254:growth & development / Q000502:physiology; D010957:Plasmids / Q000235:genetics; D017510:Protein Folding; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics
D017931:DNA Primers; D007447:Invertebrate Hormones; D011994:Recombinant Proteins; C012445:molt-inhibiting hormone