Okada Keisuke, Abe Hiroyuki, Ike Fumio, Ogura Yoshitoshi, Hayashi Tetsuya, Fukui-Miyazaki Aya, Nakamura Keiji, Shinzawa Naoaki, Horiguchi Yasuhiko
Department of Molecular Bacteriology, Research Institute for Microbial Diseases, Osaka University, Osaka, Japan.
Experimental Animal Division, RIKEN BioResource Center, Ibaraki, Japan.
PLoS One. 2015 Feb 2;10(2):e0116604. doi: 10.1371/journal.pone.0116604. eCollection 2015.
Bordetella bronchiseptica is a pathogenic bacterium causing respiratory infections in a broad range of mammals. Recently, we determined the whole genome sequence of B. bronchiseptica S798 strain isolated from a pig infected with atrophic rhinitis and found four single-nucleotide polymorphisms (SNPs) at positions -129, -72, +22, and +38 in the region upstream of dnt encoding dermonecrotic toxin (DNT), when compared with a rabbit isolate, RB50. DNT is known to be involved in turbinate atrophy observed in atrophic rhinitis. Immunoblotting, quantitative real-time PCR, and β-galactosidase reporter assay revealed that these SNPs resulted in the increased promoter activity of dnt and conferred the increased ability to produce DNT on the bacteria. Similar or identical SNPs were also found in other pig isolates kept in our laboratory, all of which produce a larger amount of DNT than RB50. Our analysis revealed that substitution of at least two of the four bases, at positions -72 and +22, influenced the promoter activity for dnt. These results imply that these SNPs are involved in the pathogenicity of bordetellae specific to pig diseases.
支气管败血波氏杆菌是一种可在多种哺乳动物中引起呼吸道感染的病原菌。最近,我们测定了从患有萎缩性鼻炎的猪身上分离出的支气管败血波氏杆菌S798菌株的全基因组序列,与兔源分离株RB50相比,发现在编码皮肤坏死毒素(DNT)的dnt基因上游区域的-129、-72、+22和+38位置有四个单核苷酸多态性(SNP)。已知DNT与萎缩性鼻炎中观察到的鼻甲萎缩有关。免疫印迹、定量实时PCR和β-半乳糖苷酶报告基因检测表明,这些SNP导致dnt启动子活性增加,并赋予细菌产生DNT的能力增强。在我们实验室保存的其他猪分离株中也发现了相似或相同的SNP,所有这些分离株产生的DNT都比RB50多。我们的分析表明,-72和+22位这四个碱基中至少两个的替换影响了dnt的启动子活性。这些结果表明,这些SNP与猪疾病特有的博德特氏菌致病性有关。