Hacker J, Schmidt G, Hughes C, Knapp S, Marget M, Goebel W
Infect Immun. 1985 Feb;47(2):434-40. doi: 10.1128/iai.47.2.434-440.1985.
The uropathogenic Escherichia coli strain 536 (O6:K15:H31) exhibits a mannose-resistant hemagglutination phenotype (Mrh) with bovine erythrocytes and delayed Mrh with human and guinea pig erythrocytes. Neuraminidase treatment of the erythrocytes abolishes mannose resistant hemagglutination, which is typical for X fimbriae. E. coli strain 536 synthesizes two different fimbriae (Fim phenotype) protein subunits, 16.5 and 22 kilodaltons in size. In addition the strain shows mannose-sensitive hemagglutination and common type I (F1) fimbriae. The cosmid clone E. coli K-12(pANN801) and another nine independently isolated Mrh+ cosmid clones derived from a cosmid gene bank of strain 536 express the 16.5-kilodalton protein band, but not the 22-kilodalton protein, indicating an association of the Mrh+ property with the "16.5-kilodalton fimbriae." All cosmid clones were fimbriated, and they reacted with antiserum produced against Mrh+ fimbriae of the E. coli strain HB101(pANN801) and lacked mannose-sensitive hemagglutination (F1) fimbriae. From the Mrh fim cosmid DNA pANN801, several subclones coding for hemagglutination and X fimbriae were constructed. Subclones that express both hemagglutination and fimbriae and subclones that only code for the hemagglutination antigen were isolated; subclones that only produce fimbriae were not detected. By transposon Tn5 mutagenesis we demonstrated that about 6.5 kilobases of DNA is required for the Mrh+ Fim+ phenotype, and the 1.5- to 2-kilobase DNA region coding for the structural protein of the fimbriae has been mapped adjacent to the region responsible for the Mrh+ phenotype. Two different regions can thus be distinguished in the adhesion determinant, one coding for hemagglutination and the other coding for fimbria formation. Transformation of plasmid DNA from these subclones into a Mrh- Fim- mutant of E. coli 536 and into a galE (rough) strain of Salmonella typhimurium yielded transformants that expressed both hemagglutination and fimbria production.
尿路致病性大肠杆菌菌株536（O6:K15:H31）对牛红细胞表现出甘露糖抗性血凝表型（Mrh），而对人及豚鼠红细胞则表现出延迟的Mrh。用神经氨酸酶处理红细胞可消除甘露糖抗性血凝，这是X菌毛的典型特征。大肠杆菌菌株536合成两种不同的菌毛（Fim表型）蛋白亚基，大小分别为16.5和22千道尔顿。此外，该菌株还表现出甘露糖敏感血凝和普通I型（F1）菌毛。黏粒克隆大肠杆菌K-12(pANN801)以及另外9个从菌株536的黏粒基因文库中独立分离得到的Mrh+黏粒克隆表达16.5千道尔顿的蛋白条带，但不表达22千道尔顿的蛋白，这表明Mrh+特性与“16.5千道尔顿菌毛”相关。所有黏粒克隆都有菌毛，它们与针对大肠杆菌菌株HB101(pANN801)的Mrh+菌毛产生的抗血清发生反应，并且缺乏甘露糖敏感血凝（F1）菌毛。从Mrh fim黏粒DNA pANN801构建了几个编码血凝和X菌毛的亚克隆。分离出了既表达血凝又表达菌毛的亚克隆以及只编码血凝抗原的亚克隆；未检测到只产生菌毛的亚克隆。通过转座子Tn5诱变，我们证明Mrh+ Fim+表型需要约6.5千碱基的DNA，并且编码菌毛结构蛋白的1.5至2千碱基DNA区域已定位在负责Mrh+表型的区域附近。因此，在黏附决定簇中可区分出两个不同区域，一个编码血凝，另一个编码菌毛形成。将这些亚克隆的质粒DNA转化到大肠杆菌536的Mrh- Fim-突变体以及鼠伤寒沙门氏菌的galE（粗糙）菌株中，得到了既表达血凝又产生菌毛的转化体。
