WSSV具蛋白酶活性肽段的检测

Identification of peptide with protease activity from white spot syndrome virus(WSSV)

在已有对虾白斑综合症病毒(WSSV)蛋白酶活性研究的基础上,从分子水平上确定WSSV呈蛋白酶活性的肽段。采用凝胶电泳技术的蛋白酶酶谱分析法确定WSSV蛋白酶活性,研究非变性条件下WSSV多肽裂解的条件,确定低温(20℃)裂解WSSV多肽可满足实验的要求。同时使用SDS-PAGE分离胶切割法结合蛋白洗脱分离出Vp19、Vp22、Vp24、Vp26和Vp28共计5个肽段,并对分离的5个组分进行蛋白酶活性的检测,初步结果表明,Vp19具有蛋白酶催化活性。

White spot syndrome virus( WSSV) is a major pathogen in the cultivated shrimp. In this research, the protein fraction of protease activity was identified. The protease activity from shrimp WSSV was charac-terized with protease profile using a combination of sodium dodecyl sulfate-polyacrylamide gel(SDS-PAGE) and protease digestion. The result showed that there was a digestion band when electrophoresis gel was in-cubated with casein after SDS-PAGE. In order to identify the peptide with protease activity, the effect of DTT and temperature for separation polypeptide of WSSV was identified. The optimum condition for split-ting polypeptide of WSSV was at low temperature(20 ℃) . The peptide component was separated by SDS-PAGE combination with electric elution. Protease activity of five elution component of WSSV (Vp19, Vp22, Vp24, Vp26, Vp28) was evaluated. The primary result was that Vp19 of WSSV is of the main component of protease-activity.