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http://dx.doi.org/10.5656/KSAE.2014.09.0.041

Inhibitory Effects of a Recombinant Viral Cystatin Protein on Insect Immune and Development  

Kim, Yeongtae (Department of Bioresource Sciences, Andong National University)
Eom, Seonghyun (Department of Bioresource Sciences, Andong National University)
Park, Jiyeong (Department of Bioresource Sciences, Andong National University)
Kim, Yonggyun (Department of Bioresource Sciences, Andong National University)
Publication Information
Korean journal of applied entomology / v.53, no.4, 2014 , pp. 331-338 More about this Journal
Abstract
Cystatins (CSTs) are reversible and competitive inhibitors of C1A cysteine proteases, corresponding to papain-like cathepsins in plants and animals. A viral CST (CpBV-CST1) was identified from a polydnavirus, Cotesia plutellae bracovirus (CpBV). Our previous study indicated that a transient expression of CpBV-CST1 interfered with immune response and development of Plutella xylostella larvae. To directly demonstrate the protein function, this study produced a recombinant CpBV-CST1 protein (rCpBV-CST1) using bacterial expression system to determine its inhibitory activity against cysteine protease and to assess its physiological alteration in insect immune and development. The open reading frame of CpBV-CST1 encodes a polypeptide of 138 amino acids (${\approx}15kDa$). rCpBV-cystatin protein in BL21 STAR (DE3) competent cells containing a recombinant pGEX4T-3:CpBV-CST1 was over-expressed by 0.5 mM IPTG for 4 h. In biological activity assay, the purified rCpBV-CST1 showed a significant inhibition against papain activity. It inhibited a cellular immune response of hemocyte nodule formation in the beet armyworm, Spodoptera exigua. Moreover, its oral administration retarded larval development of the diamondback moth, Plutella xylostella in a dose-dependent manner. These results suggest that CpBV-CST1 may be applied to control insect pest populations.
Keywords
Cystatin; Polydnavirus; Immune; Development; Recombinant protein;
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