木薯过氧化氢酶基因的克隆与原核表达

Gene Cloning and Prokaryotic Expression of Catalase in Cassava

活性氧(reactive oxygen species, ROS)的动态变化在植物的胁迫应答中起到非常重要的作用。过氧化氢酶(catalase, CAT)是动植物体内维持ROS稳态平衡的关键酶之一。为了更好地研究MeCAT1的基因特性及其在木薯响应外界环境变化的作用,本研究鉴定并克隆了木薯MeCAT1基因,成功构建MeCAT1的原核表达载体。通过蛋白结构域分析发现,MeCAT1具有过氧化氢酶的保守蛋白结构域。此后经由蛋白诱导及酶活测定试验表明MeCAT1重组蛋白具有显著的CAT酶活性。本研究为进一步探究MeCAT1在木薯应对生物胁迫和非生物胁迫过程中的作用提供了一定的分子依据,有利于MeCAT1基因在木薯育种中功能的深度挖掘和创新利用。

The dynamic changes of reactive oxygen species(ROS) play essential roles in stress response of plants.Catalase(CAT) is the one of the key enzymes regulating ROS homeostatic in animals and plants. To better investigate the genetic characteristic and the role of MeCAT1 in response to the changes of external environment in cassava, the MeCAT1 was identified and cloned, and the prokaryotic expression vector of MeCAT1 was constructed in this experiment. Protein domain analysis indicated that MeCAT1 has the conserved protein domain of catalase.Then, MeCAT1 recombinant protein showed significant CAT enzyme activity through protein induction and enzyme activity measurement. These results provides a certain molecular basis for further exploring the role of MeCAT1 in response to biotic and abiotic stresses, and it is helpful for the deep excavation and innovative utilization of MeCAT1 in cassava.