Kelly T M, Padmalayam I, Baumstark B R
Department of Biology, Georgia State University, Atlanta, Georgia 30302, USA.
Clin Diagn Lab Immunol. 1998 Nov;5(6):766-72. doi: 10.1128/CDLI.5.6.766-772.1998.
Genes coding for homologs of the highly conserved cell division protein FtsZ were isolated from Bartonella henselae and Bartonella quintana, the causative agents of cat scratch disease and trench fever, respectively. DNA fragments coding for the ftsZ open reading frames (ORFs) were cloned into Escherichia coli following PCR amplification with primers based on the ftsZ sequence of the closely related species Bartonella bacilliformis. The amino acid sequences predicted from the cloned B. henselae and B. quintana ftsZ ORFs are 81 to 83% identical to the corresponding protein in B. bacilliformis. Like the FtsZ protein of B. bacilliformis, the B. henselae and B. quintana homologs are about twice as large as the FtsZ proteins reported in most other organisms. Localized sequence differences within the C-terminal coding regions of the Bartonella ftsZ genes were used as the basis for species-specific identification of these organisms at both the DNA and protein levels. Oligonucleotide primers which permit the amplification of an ftsZ fragment from each of the Bartonella species without amplifying DNA from the other two species were designed. Anti-FtsZ antisera raised in rabbits against synthetic peptides corresponding to the relatively divergent C-terminal regions were shown via Western blot analysis to react only with the FtsZ protein from the cognate Bartonella species. These observations raise the possibility that the differences in ftsZ sequences can be used as the basis for diagnostic tests to differentiate among these closely related pathogens.
分别从猫抓病和战壕热的病原体——汉赛巴尔通体和五日热巴尔通体中分离出编码高度保守的细胞分裂蛋白FtsZ同源物的基因。基于密切相关物种杆状巴尔通体的ftsZ序列设计引物,通过PCR扩增后,将编码ftsZ开放阅读框(ORF)的DNA片段克隆到大肠杆菌中。从克隆的汉赛巴尔通体和五日热巴尔通体ftsZ ORF预测的氨基酸序列与杆状巴尔通体中的相应蛋白质有81%至83%的同一性。与杆状巴尔通体的FtsZ蛋白一样,汉赛巴尔通体和五日热巴尔通体的同源物大约是大多数其他生物体中报道的FtsZ蛋白大小的两倍。巴尔通体ftsZ基因C端编码区域内的局部序列差异被用作在DNA和蛋白质水平上对这些生物体进行物种特异性鉴定的基础。设计了寡核苷酸引物,可从每种巴尔通体物种中扩增ftsZ片段,而不会扩增其他两种物种的DNA。通过蛋白质印迹分析表明,用针对相对不同的C端区域的合成肽在兔中产生的抗FtsZ抗血清仅与同源巴尔通体物种的FtsZ蛋白发生反应。这些观察结果增加了ftsZ序列差异可作为区分这些密切相关病原体的诊断测试基础的可能性。