Chakraborty T, Huhle B, Bergbauer H, Goebel W
J Bacteriol. 1986 Jul;167(1):368-74. doi: 10.1128/jb.167.1.368-374.1986.
DNA sequences corresponding to the aerolysin gene (aer) of Aeromonas hydrophila AH2 DNA were identified by screening a cosmid gene library for hemolytic and cytotoxic activities. A plasmid containing a 5.8-kilobase EcoRI fragment of A. hydrophila DNA was required for full expression of the hemolytic and cytotoxic phenotype in Escherichia coli K-12. Deletion analysis and transposon mutagenesis allowed us to localize the gene product to 1.4 kilobases of Aeromonas DNA and define flanking DNA regions affecting aerolysin production. The reduced hemolytic activity with plasmids lacking these flanking regions is associated with a temporal delay in the appearance of hemolytic activity and is not a result of a loss of transport functions. The aerolysin gene product was detected as a 54,000-dalton protein in E. coli maxicells harboring aer plasmids and by immunoblotting E. coli whole cells carrying aer plasmids. We suggest that the gene coding aerolysin be designated aerA and that regions downstream and upstream of aerA which modulate its expression and activity be designated aerB and aerC, respectively.
通过筛选黏粒基因文库的溶血和细胞毒活性，鉴定出与嗜水气单胞菌AH2 DNA的气溶素基因（aer）相对应的DNA序列。在大肠杆菌K-12中，需要一个含有嗜水气单胞菌DNA 5.8千碱基EcoRI片段的质粒来充分表达溶血和细胞毒表型。缺失分析和转座子诱变使我们能够将基因产物定位到气单胞菌DNA的1.4千碱基，并确定影响气溶素产生的侧翼DNA区域。缺乏这些侧翼区域的质粒溶血活性降低，这与溶血活性出现的时间延迟有关，而不是运输功能丧失的结果。在携带aer质粒的大肠杆菌大细胞中，通过免疫印迹法检测携带aer质粒的大肠杆菌全细胞，气溶素基因产物被检测为一种54,000道尔顿的蛋白质。我们建议将编码气溶素的基因命名为aerA，将调节其表达和活性的aerA下游和上游区域分别命名为aerB和aerC。
