Cloeckaert A, Verger J M, Grayon M, Grépinet O
Laboratoire de Pathologie Infectieuse et Immunologie, Institut National de la Recherche Agronomique, Nouzilly, France.
Microbiology (Reading). 1995 Sep;141 ( Pt 9):2111-21. doi: 10.1099/13500872-141-9-2111.
Seventy-seven Brucella reference and field strains from different geographic origins and hosts representing the six recognized species and their different biovars were analysed for diversity of their genes encoding the major 25 and 36 kDa outer-membrane proteins (OMPs) by PCR-RFLP. The 25 kDa OMP is encoded by a single gene (omp25) whereas two closely related genes (omp2a and omp2b) encode and potentially express the 36 kDa OMP. Analysis of PCR products of the omp25 gene digested with nine restriction enzymes revealed two species-specific markers, i.e. the absence of the EcoRV site in all Brucella melitensis strains and an approximately 50 bp deletion at the 3' terminal end of the gene in all Brucella ovis strains. Analysis of PCR products of the omp2a and omp2b genes digested with 13 restriction enzymes indicated a greater diversity than the omp25 gene among the six Brucella species and within the Brucella abortus, Brucella suis, B. melitensis and B. ovis species. Greater polymorphism was also detected for the omp2b than for the omp2a gene, especially in B. ovis which seemed to carry two similar (but not identical) copies of omp2a instead of one copy each of omp2a and omp2b for the other Brucella species as was previously suggested by Ficht et al. (1990; Mol Microbiol 4, 1135-1142). Results of PCR-RFLP indicated that distinction can be made between Brucellia species and some of their biovars, except between B. canis and B. suis bv. 3 and 4, on the basis of the size and diversity of their major OMP genes, and that it could be of importance for diagnostic, epidemiological and evolutionary study purposes.
通过PCR-RFLP分析了来自不同地理来源和宿主的77株布鲁氏菌参考菌株和野外菌株，这些菌株代表了6个公认的种及其不同的生物变种，以研究其编码主要25 kDa和36 kDa外膜蛋白（OMP）的基因的多样性。25 kDa OMP由单个基因（omp25）编码，而两个密切相关的基因（omp2a和omp2b）编码并可能表达36 kDa OMP。用9种限制酶消化omp25基因的PCR产物分析显示了两个种特异性标记，即所有马尔他布鲁氏菌菌株中均不存在EcoRV位点，而所有绵羊布鲁氏菌菌株中该基因3'末端约有50 bp的缺失。用13种限制酶消化omp2a和omp2b基因的PCR产物分析表明，在6个布鲁氏菌种之间以及流产布鲁氏菌、猪布鲁氏菌、马尔他布鲁氏菌和绵羊布鲁氏菌内，其多样性比omp25基因更大。omp2b基因比omp2a基因也检测到更大的多态性，特别是在绵羊布鲁氏菌中，似乎携带两个相似（但不相同）的omp2a拷贝，而不像Ficht等人（1990年；《分子微生物学》4，1135 - 1142）之前所提出的，其他布鲁氏菌种中每个含有一个omp2a和一个omp2b拷贝。PCR-RFLP结果表明，根据其主要OMP基因的大小和多样性，可以区分布鲁氏菌种及其一些生物变种，但犬布鲁氏菌和猪布鲁氏菌生物变种３和４之间除外，并且这对于诊断、流行病学和进化研究目的可能具有重要意义。
