王先平,唐小千,周丽

• 1:中国海洋大学海水养殖教育部重点实验室

摘要(Abstract)：

为筛选一株大菱鲆致病性溶藻弧菌(Vibrio alginolyticus)SR1的热修饰性蛋白,将提取的SR1主要外膜蛋白样品与等体积电泳上样缓冲液混合,分别经28、37、50、60、70和100℃处理5min后行SDS-PAGE。发现当样品经28处理时,30kDa蛋白为外膜蛋白的主要高丰度蛋白;当样品经100℃处理时,30kDa蛋白丰度显著降低,同时出现36和37.5kDa 2条明显的蛋白条带。28℃处理样品,切胶洗脱得到的30kDa纯化蛋白,再分别经28、100℃处理后行SDS-PAGE发现只有极少量蛋白呈现30kDa,大部分呈现36和37.5kDa。切胶回收30kDa外膜蛋白制备大鼠抗血清,进行Western blotting分析。结果显示,抗血清与经各温度处理的样品中的30kDa蛋白均发生免疫交叉反应;除此之外,抗血清还与50℃处理样品中的37.5kDa蛋白及60、70和100℃处理样品中的36和37.5kDa蛋白发生免疫交叉反应。这表明30kDa外膜蛋白具有经加热处理后转化为36和37.5kDa蛋白的热修饰特性。对SR1等5株溶藻弧菌及副溶血弧菌、哈维氏弧菌等其他4种弧菌的外膜蛋白分别经28、100℃处理,SDS-PAGE分析发现5株溶藻弧菌的30kDa和副溶血弧菌的29.5kDa外膜蛋白具有热修饰性;Western blotting分析显示,抗血清可分别与28和100℃处理的5株溶藻弧菌的30kDa和副溶血弧菌的29.5kDa外膜蛋白发生免疫交叉反应,还可与100℃处理的溶藻弧菌各菌株的36kDa(以及SR1的37.5kDa)和副溶血弧菌的35kDa外膜蛋白发生免疫交叉反应。研究证实,溶藻弧菌SR1的30kDa外膜蛋白具有热修饰性,且在溶藻弧菌种内具有较高的抗原保守性。

关键词(KeyWords)： 溶藻弧菌;热修饰性外膜蛋白;免疫交叉

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(31172429);; 国家科技支撑计划项目(2012BAD17B01)资助

作者(Author): 王先平,唐小千,周丽

DOI: 10.16441/j.cnki.hdxb.2013.07.005

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