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

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• 作者：Vrinda S.^a ; ¹ ; Jasmin C.^b ; ¹ ; Sivakumar K.C^c ; ¹ ; Seena Jose^a ; ¹ ; Blessy Jose^a ; ¹ ; Rosamma Philip^d ; ¹ ; Bright Singh I. S.^a ; ¹ ; ^{isbsingh@gmail.com}
• 关键词：Gonad-inhibiting hormone (GIH) ; Fusion/chimeric protein ; Thioredoxin-fused protein ; Penaeus monodon ; CHH family hormone ; XO-SG complex ; GIH polyclonal antisera
• 刊名：Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology
• 出版年：2017
• 出版时间：January 2017
• 年：2017
• 卷：203
• 期：Complete
• 页码：167-178
• 全文大小：2794 K
• 卷排序：203

文摘

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.