Expression, Purification And Polyclonal Antibody Preparation Of Molt-inhibiting Hormone 1 From Chinese Mitten Crab Eriocheir Japonica Sinensis

Crustaceans grow by periodic molting. Molting in Crustaceans is controlled by the molt-inhibiting hormone (MIH) and ecdysone. MIH negatively regulates the synthesis of ecdysteroid molting hormones produced by crustacean Y-organs, resulting in molt suppression. It is a neuropeptide member belonging to the crustacean hyperglycemic hormone (CHH) neuropeptide family. Besides MIH, CHH family neuropeptides include the crustacean hyperglycemic hormone (CHH), gonad-inhibiting hormone (GIH) and mandibular organ-inhibiting hormone (MOIH). These peptides have common amino acid compositions and organizations. All members of this family generally have molecular weights of 7-9 kD consisted of approximately 70-78 amino acids and three disulfide bridges. These neuropeptides, produced in the medulla terminalis of the X-organ (MTXO) and stored in the sinus gland of the eyestalk, played an important role in the blood glucose regulation and regulated several important physiological processes including growth and reproduction.Although recent progress has been made in understanding the structure and function of MIH, important questions remain unanswered. The detail mechanism of MIH actions is unknown, and the expression of MIH during development has not been investigated. In the present study, molt-inhibiting hormone-1 from Eriocheir japonica sinensis (Ers-MIH1) was studied both in prokaryotic and eukaryotic expression system. The resulting MIH1fused protein was purified, and used as immunogen to prepare the polyclonal antibody.