Donnenberg M S, Tzipori S, McKee M L, O'Brien A D, Alroy J, Kaper J B
Medical Service, Department of Veterans Affairs Medical Center, Baltimore, Maryland.
J Clin Invest. 1993 Sep;92(3):1418-24. doi: 10.1172/JCI116718.
The eaeA gene of enteropathogenic Escherichia coli (EPEC) is necessary for intimate attachment to epithelial cells in vitro. Enterohemorrhagic E. coli (EHEC) strains also possess an eae gene and are capable of intimate attachment and microvillus effacement in vitro and in animal models. To assess the role of the EHEC eae gene in intimate attachment, we constructed an eae deletion/insertion mutation in wild-type EHEC O157:H7 strain 86-24 by using linear electroporation of a recombinant allele. The mutant obtained was deficient in inducing f-actin accumulation in HEp-2 cells and was incapable of attaching intimately to colonic epithelial cells in a newborn piglet model of infection. Intimate attachment in vivo was restored when the EHEC eae gene or the eaeA gene of EPEC was introduced into the mutant on a plasmid. These results indicate that the eae gene is necessary for intimate attachment of EHEC in vivo. In addition, the complementation achieved by the EPEC locus indicates that the eae gene of EHEC and the eaeA gene of EPEC are functionally homologous.
肠致病性大肠杆菌（EPEC）的eaeA基因对于其在体外紧密黏附上皮细胞是必需的。肠出血性大肠杆菌（EHEC）菌株也拥有一个eae基因，并且能够在体外和动物模型中进行紧密黏附和微绒毛缺失。为了评估EHEC的eae基因在紧密黏附中的作用，我们通过重组等位基因的线性电穿孔在野生型EHEC O157:H7菌株86 - 24中构建了一个eae缺失/插入突变体。获得的突变体在诱导HEp - 2细胞中f - 肌动蛋白积累方面存在缺陷，并且在新生仔猪感染模型中无法紧密黏附于结肠上皮细胞。当将EHEC的eae基因或EPEC的eaeA基因通过质粒导入突变体时，体内的紧密黏附得以恢复。这些结果表明，eae基因对于EHEC在体内的紧密黏附是必需的。此外，由EPEC位点实现的互补表明，EHEC的eae基因和EPEC的eaeA基因在功能上是同源的。
