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    Regulating gonad inhibition and vitellogenin/vitellin induction in Penaeus monodon using mature GIH fusion protein and polyclonal antisera

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    Comp_Biochem_Physiol_203_167.pdf (9.374Mb)
    Date
    2017
    Author
    Vrinda, S.
    Jasmin, C.
    Sivakumar, K.C.
    Jose, S.
    Jose, B.
    Philip, R.
    BrightSingh, I.S.
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    Abstract
    Gonad inhibiting hormone (GIH), type II class of the CHH family neuropeptides, is released by the neurohaemal XO-SG complex of the eyestalk. The inhibitory function of GIH has a pivotal role in gonad development and reproduction. In this study, we report the expression and production of a thioredoxin-fused mature GIH protein (mf-PmGIH) of Penaeus monodon in a bacterial system and its use as antigen to raise polyclonal antiserum (anti-mf-PmGIH). The mature GIH gene of 237 bp that codes for 79 amino acids, was cloned into the Escherichia coli thioredoxin gene fusion expression system. The expression vector construct (mf-PmGIH + pEt32a +) upon induction produced 32.16 kDa mature GIH fusion protein (mf-PmGIH)·The purified fusion protein was used as exogenous GIH and as antigen to raise polyclonal antisera. The fusion protein when injected into juvenile shrimp significantly reduced vitellogenin/vitellin levels by 31.55% within 72 h in comparison to the controls showing the gonad inhibiting property. Vitellogenin/vitellin levels were significantly induced by 74.10% within 6 h when polyclonal antiserum (anti-mf-PmGIH-1:500) was injected in P.monodon. Anti-mf-PmGIH immunolocalized GIH producing neurosecretory cells in the eyestalk of P.monodon. The present manuscript reports an innovative means of gonad inhibition and vitellogenin/vitellin induction with thioredoxin fused GIH and antisera developed
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    http://drs.nio.org/drs/handle/2264/5076
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