title: Moult-inhibiting fusion protein augments while polyclonal antisera attenuate moult stages and duration in Penaeus monodon.
abstract: Moulting in crustaceans is regulated by moult-inhibiting hormone (MIH) of the CHH family neuropeptides. The inhibitory functions of MIH have pivotal roles in growth and reproduction of Penaeus monodon. In this study, we report the expression of a thioredoxin-fused mature MIH I protein (mf-PmMIH I) of P. monodon in a bacterial system and its use as antigen to raise polyclonal antiserum (anti-mf-PmMIH I). The mature MIH I gene of 231bp, that codes for 77 amino acids, was cloned into the Escherichia coli thioredoxin gene fusion expression system. The translation expression vector construct (mf-PmMIH I+pET32a+) upon induction produced 29.85kDa mature MIH I fusion protein (mf-PmMIH I). The purified fusion protein was used as exogenous MIH I and as antigen to raise polyclonal antisera. When fusion protein (mf-PmMIH I) was injected into D2 and D3 stages of juvenile shrimp, the moult cycle duration was extended significantly to 16.67±1.03 and 14.67±1.03days respectively compared to that of 11.67±1.03days in controls. Moult duration was further reduced to 8.33±0.82days when polyclonal antiserum (anti-mf-PmMIH I - 1:500 dilutions) was injected. Anti-mf-PmMIH I immunolocalized MIH I producing neurosecretory cells in the eyestalk of P. monodon. In short, the present manuscript reports an innovative means of moult regulation in P. monodon with thioredoxin fused MIH I and antisera developed.
authors.affiliation: National Centre for Aquatic Animal Health, Cochin University of Science and Technology, Kochi 16, Kerala, India.
authors.forename: S
authors.initials: S
authors.lastname: Vrinda
authors.affiliation: CSIR - National Institute of Oceanography, Regional Centre, Kochi 18, Kerala, India.
authors.forename: C
authors.initials: C
authors.lastname: Jasmin
authors.affiliation: Bioinformatics Facility, Rajiv Gandhi Centre for Biotechnology, Thiruvananthapuram 14, Kerala, India.
authors.forename: K C
authors.initials: KC
authors.lastname: Sivakumar
authors.affiliation: National Centre for Aquatic Animal Health, Cochin University of Science and Technology, Kochi 16, Kerala, India.
authors.forename: Blessy
authors.initials: B
authors.lastname: Jose
authors.affiliation: Department of Marine Biology, Microbiology and Biochemistry, Cochin University of Science and Technology, Kochi 16, Kerala, India.
authors.forename: Rosamma
authors.initials: R
authors.lastname: Philip
authors.affiliation: National Centre for Aquatic Animal Health, Cochin University of Science and Technology, Kochi 16, Kerala, India. Electronic address: isbsingh@gmail.com.
authors.forename: I S
authors.initials: IS
authors.lastname: Bright Singh
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000494:pharmacology*; D005260:Female; D007106:Immune Sera / Q000494:pharmacology; D007447:Invertebrate Hormones / Q000235:genetics / Q000276:immunology / Q000378:metabolism / Q000494:pharmacology*; D051379:Mice; D008807:Mice, Inbred BALB C; D018978:Molting / Q000187:drug effects* / Q000502:physiology; D033561:Penaeidae / Q000187:drug effects / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D013879:Thioredoxins / Q000235:genetics / Q000276:immunology / Q000378:metabolism / Q000494:pharmacology*; D013997:Time Factors
chemical_list: D000906:Antibodies; D007106:Immune Sera; D007447:Invertebrate Hormones; D011993:Recombinant Fusion Proteins; C012445:molt-inhibiting hormone; D013879:Thioredoxins
keywords: Fusion recombinant protein; MIH polyclonal antibody; Moult-inhibiting hormone (MIH); Penaeus monodon; XO-SG complex