Welch R A
Infect Immun. 1987 Sep;55(9):2183-90. doi: 10.1128/iai.55.9.2183-2190.1987.
DNA sequences similar to those of the Escherichia coli hemolysin genes were detected among uropathogenic isolates of Proteus vulgaris and Morganella morganii by using the Southern blotting technique and hly gene-specific DNA probe. Immunoblotting revealed that among the hemolytic P. vulgaris and M. morganii isolates there was expressed a polypeptide species similar in molecular size (110 kilodaltons) and antigenicity to Escherichia coli HlyA. A plasmid-mediated P. vulgaris hemolysin determinant identified by Southern blotting analysis was molecularly cloned, and the recombinant plasmid (pWPV100) was characterized by restriction endonuclease fragment mapping. A second recombinant library of genomic DNA prepared from a hemolytic, urinary tract isolate of Proteus mirabilis was constructed in E. coli. A 5.5-kilobase XhoI fragment encoding an extracellular hemolytic activity was molecularly cloned (pWPM100), and this plasmid was subjected to transposon-mediated mutagenesis with TnphoA. The P. mirabilis hemolytic phenotype was determined to be encoded by a polypeptide species (HpmA) with an estimated molecular size of 140 kilodaltons based on minicell polypeptide analysis of pWPM100 and its mutant derivatives. Southern blotting analysis with a HpmA-specific DNA probe revealed that this novel determinant is commonly found in both Proteus species but is not present in hemolytic isolates of M. morganii, E. coli, Citrobacter freundii, and Serratia marcescens.
利用Southern印迹技术和hly基因特异性DNA探针，在普通变形杆菌和摩根摩根菌的尿路致病性分离株中检测到了与大肠杆菌溶血素基因序列相似的DNA序列。免疫印迹显示，在溶血的普通变形杆菌和摩根摩根菌分离株中表达了一种分子大小（110千道尔顿）和抗原性与大肠杆菌HlyA相似的多肽。通过Southern印迹分析鉴定的质粒介导的普通变形杆菌溶血素决定簇进行了分子克隆，重组质粒（pWPV100）通过限制性内切酶片段图谱进行了表征。从奇异变形杆菌的溶血尿路分离株制备的基因组DNA的第二个重组文库在大肠杆菌中构建。编码细胞外溶血活性的5.5千碱基XhoI片段进行了分子克隆（pWPM100），该质粒用TnphoA进行转座子介导的诱变。基于对pWPM100及其突变衍生物的小细胞多肽分析，确定奇异变形杆菌的溶血表型由一种估计分子大小为140千道尔顿的多肽（HpmA）编码。用HpmA特异性DNA探针进行的Southern印迹分析表明，这种新的决定簇在两种变形杆菌中普遍存在，但在摩根摩根菌、大肠杆菌、弗氏柠檬酸杆菌和粘质沙雷氏菌的溶血分离株中不存在。
