摘要
目的 了解约氏疟原虫感染雌斯氏按蚊成蚊是否发生黑化相关性酶级联反应。方法 挤压法收集雌斯氏按蚊成蚊血淋巴 ,Bradford法检测血淋巴蛋白浓度 ,然后对血淋巴进行一维和二维SDS PAGE ,继而利用Western印迹分析比较PPO蛋白点的差异。结果 约氏疟原虫感染雌斯氏按蚊成蚊之前的血淋巴蛋白二维凝胶电泳后的Western印迹的PVDF膜上出现 2个分子量均约为 64× 10 3、等电点分别约为 7 4和 7 6的两个PPO蛋白点 ,感染后发现 8个蛋白点 ,3 6×10 3分子量处 3个 ,等电点分别约为 7 2、7 4和 8 0 ;3 0× 10 3分子量处 5个 ,等电点分别约为 7 0、7 2、7 5、7 6和 7 8。结论 约氏疟原虫感染雌斯氏按蚊成蚊之前的血淋巴PPO以异二聚体形式结构性表达于血淋巴内 ,感染后PPO发生免疫激活反应和酶学分解过程 ,提示PPO参与疟原虫卵囊黑化包被反应。
Objective To explore occurrence or not of the cascade reaction related with melanization in Anopheles stephensi adult females infected with Plosmodium yoelii . Methods The hemolymph of Anopheles stephensi adult females was collected by expulsion method. The concentration of hemolymph protein was examined by Bradford method, and then the hemolymph was analyzed by two dimensional electrophoresis (2 DE) and SDS polyacrylamide gel electrophoresis (SDS PAGE). The differential PPO spots were identified and compared by Western blotting. Results Western blot analysis after 2 DE revealed that there were two specifically separated PPO protein spots (molecular weight: 64×10 3, isoelectric points: 7.4 and 7.6) in the polyvinyl difluoride membrane before Plosmodium yoelii infection. After infection, however, there were 8 PPO spots (3 spots with molecular weight of 36×10 3, isoelectric points at 7.2, 7.4, and 8.0; 3 spots with molecular weight of 30×10 3, isoelectric points at 7.0, 7.2, 7.5, 7.6, and 7.8). Conclusion PPO is constitutively expressed in the hemolymph of Anopheles stephensi adult females and exists as heterodimers in vivo before infection. After Plosmodium yoelii infection, however, immunological activation response and zymolysis of PPO in the hemolymph of Anopheles stephensi suggests that PPO participates in the melanotic encapsulation of oocysts of Plosmodium yoelii .
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2004年第6期506-508,共3页
Journal of Third Military Medical University
基金
国家自然科学基金资助项目 ( 30 30 0 2 92 )~~
关键词
斯氏按蚊
约氏疟原虫
血淋巴
前酚氧化酶
双向电泳
WESTERN印迹
Anopheles stephensi
Plosmodium yoelii
prophenoloxidase
hemolymph
two dimensional gel electrophoresis
Western blotting
