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【目的】研究溶藻弧菌的溶血现象,溶血素基因vah的分布及vah基因、vah启动子区对溶藻弧菌溶血活性的贡献。【方法】对46株分离自华南沿海水生动物体内和海水的溶藻弧菌环境株及溶藻弧菌标准株1.1587进行溶血实验;比较具有溶血活性的溶藻弧菌野生株ZJ051、vah基因大肠杆菌BL21重组表达株、vah缺失突变株和基因回补株间溶血能力的差异;检测vah基因在溶藻弧菌中的分布,比较溶血株与非溶血株vah基因及上游启动子区的序列差异。【结果】47.8%的溶藻弧菌菌株产生溶血活性,因此溶血现象普遍存在于溶藻弧菌环境株中;vah基因的表达产物具有溶血活性,vah基因缺失突变株不具有溶血活性,而vah基因回补株恢复溶血活性。vah基因普遍存在于溶藻弧菌中,且基因序列非常相似,氨基酸序列完全相同,然而不同菌株的启动子区第188?190碱基位点存在差异。【结论】溶藻弧菌vah基因是造成溶藻弧菌溶血的直接原因,但溶藻弧菌溶血能力的差异并非是由vah基因本身差异决定,极有可能与启动子区第188?190碱基位点相关。
【Objective】 The purpose of this study was to investigate the hemolysis of Vibrio alginolyticus, the distribution of the hemolysin gene vah and the contribution of the vah promoter and vah promoter to the hemolytic activity of V. alginolyticus. 【Method】 46 strains of V. alginolyticus isolated from the coastal animals in South China Sea and seawater were tested for hemolytic activity with Vibrio alginolyticus 1.1587. The hemolytic activity of Vibrio alginolyticus ZJ051, Bacillus BL21 recombinant expression strain, vah deletion mutant and gene complement strains hemolytic ability differences; vah gene distribution in Vibrio alginolyticus distribution, comparison of hemolysin and non-hemolytic strains vah gene and upstream promoter region of the sequence difference . 【Results】 47.8% strains of V. alginolyticus produced hemolytic activity. Therefore, hemolysis was common in V. alginolyticus strains. The expression product of vah gene was hemololytic and the deletion mutant of vah gene did not have hemolytic activity. Gene replenishing strains restore hemolytic activity. The vah gene is commonly found in Vibrio alginolyticus and has very similar gene sequences and identical amino acid sequences. However, the 188th to 190th bases in the promoter region of different strains are different. 【Conclusion】 V. alginolyticus vah gene is the direct cause of hemolysis of V. alginolyticus, but the difference of hemolysis ability of V. alginolyticus is not determined by the difference of vah gene itself, Base point related.