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

바이러스 유래 시스타틴 재조합 단백질의 곤충 면역 및 발육 억제효과

  • 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)
  • 김영태 (안동대학교 생명자원과학과) ;
  • 엄성현 (안동대학교 생명자원과학과) ;
  • 박지영 (안동대학교 생명자원과학과) ;
  • 김용균 (안동대학교 생명자원과학과)
  • Received : 2014.07.18
  • Accepted : 2014.09.18
  • Published : 2014.12.01

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.

Acknowledgement

Supported by : 농촌진흥청

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