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对六株嗜水气单胞菌的全细胞和外膜成份在不同的培养条件下进行SDS-PAGE、免疫转印分析.在同一培养条件下6个供试菌株全细胞的蛋白质电泳图谱不同.同一菌株在五种不同的培养条件下,其蛋白质电泳图谱相似但有差异.6株菌株的细菌外膜提取物的蛋白质电泳图谱不同,同一菌株在5种不同的培养条件下,其蛋白质电泳图谱相似但有差异,但所有的图谱均具有34 kDa和30 kDa的条带.用Schiff s试剂,银染及生物素标记(BS-1,ConA,UEA-1)法测定及分析提取物中糖或碳水化合物的特征.生物素标记及经蛋白酶K处理后的所有菌株的提取物均显示不同分子量碳水化合物的条带,但具有相同分子量的条带,其中32 kDa条带被三种生物素及银染所显示,而34 kDa仅被生物素BS-1和UEA-1所识别.经Schiff s试剂染色后,仅出现少量的条带.用免疫转印法测定菌体提取物的抗原性,结果显示,提取物有多条不同的条带被两种血清所识别,且用活菌制备的血清比死菌的要强得多.所有菌株在不同培养条件下的提取物均具35 kDa条带被鱼血清所识别.所以,35 kDa条带可能是嗜水气单胞菌的共同抗原.同时,用ELISA法测定,证明鱼血清与其它菌株间有较强的交叉反应.
The whole cell and adventitial components of six Aeromonas hydrophila strains were analyzed by SDS-PAGE and immunoblot under different culture conditions.The protein profiles of the whole cell of the six tested strains were different under the same culture conditions. The same strains under the five different culture conditions, the protein electrophoresis patterns are similar but there are differences.The six strains of bacterial outer membrane extracts of different protein electrophoresis patterns, the same strain in five different culture conditions, the protein electrophoresis pattern Similar but different, but all of the bands had bands of 34 kDa and 30 kDa. The amount of sugar in the extract was determined and analyzed using Schiff’s reagent, silver stain and biotinylated (BS-1, ConA, UEA-1) Or carbohydrates.Biotin labeling and extracts of all strains after protease K treatment showed bands of different molecular weight carbohydrates but bands of the same molecular weight with a 32 kDa band surrounded by three biotin and Silver staining, 34 kDa was only recognized by biotin BS-1 and UEA-1, only a few bands appeared after staining with Schiff’s reagent.The antigenicity of the bacterial extract was determined by immunotransfer method, The results showed that there are a number of extracts do not Of the bands were identified by both sera and the serum prepared with viable bacteria was much more potent than the dead bacteria.The extracts of all strains under different culture conditions were identified by the fish serum as 35 kDa bands kDa band may be the common antigen of Aeromonas hydrophila.At the same time, the results of ELISA showed that there was a strong cross-reaction between fish serum and other strains.