Bahrani F K, Cook S, Hull R A, Massad G, Mobley H L
Division of Infectious Diseases, University of Maryland School of Medicine, Baltimore, Maryland 21201.
Infect Immun. 1993 Mar;61(3):884-91. doi: 10.1128/iai.61.3.884-891.1993.
Proteus mirabilis, a common cause of urinary tract infection in hospitalized and catheterized patients, produces mannose-resistant/klebsiella-like (MR/K) and mannose-resistant/proteus-like (MR/P) hemagglutinins. The gene encoding the major structural subunit of a fimbria, possibly MR/K, was identified in two strains. A degenerate oligonucleotide probe based on the N terminus of the Proteus uroepithelial cell adhesin and antiserum raised against the denatured polypeptide were used to screen a cosmid gene bank of strain HU1069. A cosmid clone that reacted with the probe and antiserum was identified, and a fimbria-like open reading frame was determined by nucleotide sequencing. The predicted N-terminal amino acid sequence of the processed polypeptide, ENETPAPKVSSTKGEIQLKG (residues 23 to 42), did not match the uroepithelial cell adhesin N terminus but, rather, matched exactly the N-terminal amino acid sequence of a polypeptide with an apparent molecular size of 19.5 kDa isolated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of a fimbrial preparation from strain HI4320 expressing MR/K hemagglutinin. By using an oligonucleotide from the HU1069 open reading frame, the fimbrial gene was isolated and sequenced from a cosmid gene bank clone of strain HI4320. A 552-bp open reading frame predicts a 184-amino-acid polypeptide including a 22-amino-acid hydrophobic leader sequence. The unprocessed polypeptide is predicted to be 18,921 Da; the processed polypeptide is predicted to be 16,749 Da. The predicted amino acid sequence of the polypeptide encoded by the gene, designated pmfA, displayed 36% exact matches with the mannose-resistant fimbrial subunit encoded by smfA of Serratia marcescens but only 15% exact matches with the predicted sequence encoded by mrkA of Klebsiella pneumoniae.
奇异变形杆菌是住院患者和留置导尿管患者尿路感染的常见病因,可产生抗甘露糖/肺炎克雷伯菌样(MR/K)和抗甘露糖/变形杆菌样(MR/P)血凝素。在两株菌株中鉴定出了编码一种菌毛主要结构亚基(可能是MR/K)的基因。基于变形杆菌尿道上皮细胞粘附素N端的简并寡核苷酸探针和针对变性多肽产生的抗血清被用于筛选菌株HU1069的黏粒基因文库。鉴定出了一个与探针和抗血清发生反应的黏粒克隆,并通过核苷酸测序确定了一个类似菌毛的开放阅读框。加工后多肽的预测N端氨基酸序列ENETPAPKVSSTKGEIQLKG(第23至42位氨基酸残基)与尿道上皮细胞粘附素的N端不匹配,而是与通过对表达MR/K血凝素的菌株HI4320的菌毛制剂进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分离得到的表观分子大小为19.5 kDa的多肽的N端氨基酸序列完全匹配。通过使用来自HU1069开放阅读框的寡核苷酸,从菌株HI4320的黏粒基因文库克隆中分离并测序了菌毛基因。一个552 bp的开放阅读框预测了一个184个氨基酸的多肽,包括一个22个氨基酸的疏水前导序列。未加工的多肽预计为18,921 Da;加工后的多肽预计为16,749 Da。该基因编码的多肽(命名为pmfA)的预测氨基酸序列与粘质沙雷氏菌smfA编码的抗甘露糖菌毛亚基有36%的完全匹配,但与肺炎克雷伯菌mrkA编码的预测序列只有15%的完全匹配。