中文摘要：

为筛选一株大菱鲆致病性溶藻弧菌（Vibrio alginolyticus）SRl的热修饰性蛋白，将提取的SRl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37．5kDa2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。

英文摘要：

Major outer membrane proteins （MOMPs） of a pathogenic Vibrio alginolyticus SR1 were prepared in order to select the heat-modifiable proteins. The MOMPs were mixed with an equal volume of loading buffer, heated at 28, 37, 50, 60, 70 and 100℃, respectively, and separated with sodium dodecyl sulfate-polyacrylamide gel electrophoresis （SDS-PAGE）. A major outer membrane protein （MOMP）, 30 kDa in molecular weight, was the only highly abundant protein when the MOMPs were heated at 28℃, which turned to be a less abudant protein when MOMPs were heated at 100℃. Two new proteins, 36 kDa and 37. 5 kDa in molecular weight, appeared as the abundant proteins when MOMPs were heated at 100 ~C, which may derived from the protein of 30 kDa. Mouse antiserum against the protein of 30 kDa recovered from gel recognized proteins of 30 kDa, 36 kDa and 37.5 kDa on the Western blot of MOMPs heated at different temperatures, indicating that 30 kDa MOMP was heat modifiable. The antiserum recognized also the protein of 30 kDa of 5 strains of V. alginolyticus and the protein of 29.5 kDa of V. parahaemolyticus when their MOMPs were heated at 28 and 100℃, respectively. The recognized included also 36 kDa of 5 strains of V. alginolyticus （also including the 37.5 kDa of SR1）and 35 kDa of V. parahaemolyticuswhen their MOPMs were heated at 100 ℃These results proved that the protein of 30 kDa of V. alginolyticus SR1 was a major heat-modifiable protein, which was highly conservative in antigenicity among different strains of V. alginolyticus.