Expression and characterization of recombinant Juvenile Hormone Epoxide Hydrolase of Spilarctia obliqua, a major pest in Agri-Sericulture
Swetha Kumari K1
Mamatha Dadala Mary2
1. Research Fellows, Dept. of Sericulture, Sri Padmavati Women’s University, Tirupati, India
2. Associate Professor, Dept. of Sericulture, Sri Padmavati Women’s University, Tirupati, India
3. Intern Student, University Preparatory Academy, San Jose, California, United States of America
Molecular Entomology, 2015, Vol. 6, No. 1 doi: 10.5376/me.2015.06.0001
Received: 27 Aug., 2014 Accepted: 13 Oct., 2014 Published: 01 Jan., 2015
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Preferred citation for this article:
Swetha kumari et al., 2015, Expression and characterization of recombinant Juvenile Hormone Epoxide Hydrolase of Spilarctia obliqua, a major pest in Agri-Sericulture, Molecular Entomology, Vol.6, No.1 1-11 (doi: 10.5376/me.2015.06.0001)
Juvenile Hormone (JH) modulates a variety of developmental and physiological processes in Insects. Previous studies suggest that the two hydrolytic enzymes namely Juvenile Hormone Esterase (JHE) and Juvenile Hormone Epoxide Hydrolase (JHEH) play important roles in the regulation of JH titer. Compared with JHE, JHEH is more essential regulatory enzyme and could be applied to control insects, because of its irreversible reaction on JH Mechanism. A full length of cDNA (1323bp) encoding juvenile hormone epoxide hydrolase gene from Spilarctia obliqua (SoJHEH) is isolated and subcloned into baculovirus vector system using pFastBac and pFastBac-HTA vectors. The bacmid-transformed SoJHEH is expressed in Insect cell line system and the expression of recombinant SoJHEH is confirmed by Western blotting. The enzyme activity of recombinant SoJHEH showed a high activity with JH III and very low with other general epoxide substrates. The deduced SoJHEH protein sequence showed the conserved domains of catalytic triad and oxyanion hole indicating the well preserved catalytic mechanism of epoxide hydration which can be explored further for bio control strategy at molecular level using insect’s own enzyme which can address the current problem of insect resistance.
Spilarctia obliqua; Juvenile Hormone; Juvenile hormone epoxide Hydrolase; Baculovirus expression system; Characterization