Benz I, Schmidt M A
Zentrum für Molekulare Biologie Heidelberg, Heidelberg, Germany.
Res Microbiol. 1990 Sep-Oct;141(7-8):785-6. doi: 10.1016/0923-2508(90)90111-3.
For the identification and characterization of the factor(s) responsible for the diffuse adherence (DA) pattern of enteropathogenic Escherichia coli strains, E. coli strain 2787 isolated from a case of infantile diarrhoea was employed. A plasmid-derived 11-kb fragment was cloned into pBR322. The recombinant plasmid pIB6 was shown to confer the diffuse adherence phenotype on different E. coli K12 strains as well as pIB4, a plasmid with a 9.2-kb insert. The DNA fragment necessary for the expression of the DA phenotype could be reduced to 6.0 kb. Antiserum obtained against pIB4-encoded proteins recognized a surface-associated protein of about 100 kDa in Western blotting. The isolated 100-kDa protein was found to bind to HeLa cells. The antiserum against C600(pIB4) inhibits adherence of E. coli 2787 and C600(pIB6) to HeLa cells. For this reason, the protein is called adhesin involved in diffuse adherence (AIDA-I).
为了鉴定和表征导致致病性大肠杆菌菌株弥散黏附(DA)模式的因子,采用了从一例婴儿腹泻病例中分离出的大肠杆菌菌株2787。将一个源自质粒的11kb片段克隆到pBR322中。重组质粒pIB6以及带有9.2kb插入片段的质粒pIB4,被证明可赋予不同的大肠杆菌K12菌株弥散黏附表型。表达DA表型所需的DNA片段可缩减至6.0kb。针对pIB4编码蛋白获得的抗血清在蛋白质印迹法中识别出一种约100kDa的表面相关蛋白。发现分离出的100kDa蛋白可与HeLa细胞结合。针对C600(pIB4)的抗血清可抑制大肠杆菌2787和C600(pIB6)对HeLa细胞的黏附。因此,该蛋白被称为参与弥散黏附的黏附素(AIDA-I)。