嗜水气单胞菌322A的外膜蛋白及其抗原性

Antigenicity of outer membrane proteins of Aeromonas hydrophila

用十二烷基肌氨酸钠抽提结合超速离心的方法提取一株日本鳗鲡致病性嗜水气单胞菌322A和其他4株气单胞菌标准菌株的外膜蛋白(Omp);通过SDS-PAGE分析比较这5株气单胞菌Omp的组成.结果表明,5株气单胞菌的Omp电泳可得到5-13条条带,其分子质量主要集中在20-50 ku之间,且28 ku的Omp为5株气单胞菌所共有.用罗非鱼感染322A全菌后的恢复期血清进行蛋白质印迹试验的结果显示,322A的Omp条带中有4条发生阳性反应,其分子质量分别为28、37、43和45 ku.用兔抗322A Omp的血清进行蛋白质印迹试验的结果显示,322A的Omp条带中有5条发生阳性反应,其分子质量分别为28、33、37、43和45 ku.不同抗血清的印迹结果提示了322A数条主要Omp具有免疫原性,有望成为疫苗研发的候选材料.

A pathogenic Aeromonas hydrophila strain 322A was isolated from Anguillajaponica.Strain 322A and other four strains of Aeromonas were used for preparation of the outer membrane proteins （Omps） with Sarkosyl method combining super centrifugal purification.SDS-PAGE showed that there were generally 5-13 Omp bands in each of the 5 strains.Molecular weight focused on 20-50 ku,and the protein with molecular weight of 28 ku was common for all the 5 strains.After SDS-PAGE,in the Western-blot with tilapia convalescent antiserum against 322A,only 4 bands of strain 322A with molecular weight of 28,37,43 and 45 ku were detected.In the Western-blot with rabbit antiserum against Omps of 322A,5 bands of strain 322A with molecular weight of 28,33,37,43 and 45 ku were detected.The results of Western-blot suggested that several major outer membrane proteins of strain 322A had immunogenicity,which may be vaccine candidates.