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Thioredoxin fused molt-inhibiting hormone and polyclonal anti-sera: Effect on molt duration in Penaeus monodon
6th World Congress on Biotechnology
Vrinda S, Jasmine C, Seena Jose, Blessy Jose, Rosamma Philip and Bright Singh
Cochin University of Science and Technology, India
ScientificTracks Abstracts: J Biotechnol Biomater
Abstract
The inhibitory functions of Moult Inhibiting Hormone (MIH) have a pivotal role in growth and reproduction of Penaeus monodon. In this study we report the production of biologically active thioredoxin-fused mature MIH I protein (mf- PmMIH I) of P. monodon for the first time in a bacterial system and its use as an antigen to raise polyclonal anti-sera (anti-mf- PmMIH I). Using reverse transcription, the open reading frame (ORF) of 357 bp cDNA sequence of MIH I was isolated from the eyestalk of P. monodon cultured in the Indian brackish water rearing culture system. The mature MIH I gene of 231 bp that codes for 77 amino acids was cloned into the Escherichia coli thioredoxin gene fusion expression system. Upon induction 29.85 kDa mature MIH I fusion protein (mf-PmMIH I) was expressed, purified and refolded for use as exogenous MIH I and as an antigen to raise polyclonal antisera. The specificity and sensitivity of the raised polyclonal antisera against the mf-PmMIH I antigen was examined by Western blot, ELISA and immunofluorescence. The thioredoxin-fused mature MIH I protein and the polyclonal antisera developed were tested in adult P. monodon (n=20, 10-15 g) to determine their acute biological effects on augmentation and attenuation of molt cycle duration. In the shrimps injected with mf-PmMIH I, the molt cycle duration was significantly increased to 16.67�?±1.03 days while the anti-mf-PmMIH I exhibited a molt duration that was significantly reduced to 8.33�?±0.82 days. This innovative MIHI fusion protein appears to be practical in terms of effective molting.Biography
Vrinda S has completed her PhD in Marine Biotechnology on the topic “Development of CHH family recombinant hormones and RNAi for induced maturation of Penaeus monodon” from Cochin University of Science and Technology, Kerala. She is a Dr. D S Kothari Postdoctoral Fellow pursuing her Postdoctoral studies at National Centre for Aquatic Animal Health, CUSAT on the topic entitled “Recombinant antimicrobial peptides and cell penetrating domains from Phylum Arthropod and Mollusc as anticancer agents”.
Email: vrinda.sukumaran@gmail.com
