Bardarov S S, Sirakova T D, Kriakov J, Karakashyan A, Markov K
Department of Microbiology, Bulgarian Academy of Sciences, Sofia.
FEMS Microbiol Lett. 1990 Sep 15;59(3):277-9. doi: 10.1016/0378-1097(90)90233-g.
Chromosomal DNA from reference Yersinia strains was digested individually with 9 restriction endonucleases. DNA fragments were separated and analyzed by electrophoresis through agarose gels. The clearest fragment patterns were obtained when EcoRI was employed. The Y. pestis fragment pattern obtained after the use of this enzyme showed the presence of a unique DNA fragment with molecular mass 1400 bp. This DNA fragment was cloned, purified, labeled with 32P and then used to probe EcoRI digests of all three Yersinia species. A strong hybridization signal was obtained with Y. pestis strain. No such signal was found with Y. pseudotuberculosis or Y. enterocolitica. These results indicate that the DNA fragment is species specific and could be used as a diagnostic DNA probe for Y. pestis.
用9种限制性内切核酸酶分别消化参考耶尔森氏菌菌株的染色体DNA。通过琼脂糖凝胶电泳分离并分析DNA片段。使用EcoRI时获得的片段模式最清晰。使用该酶后获得的鼠疫耶尔森氏菌片段模式显示存在一个分子量为1400 bp的独特DNA片段。该DNA片段被克隆、纯化，用32P标记，然后用于探测所有三种耶尔森氏菌的EcoRI消化产物。鼠疫耶尔森氏菌菌株获得了强烈的杂交信号。在假结核耶尔森氏菌或小肠结肠炎耶尔森氏菌中未发现此类信号。这些结果表明该DNA片段具有种特异性，可作为鼠疫耶尔森氏菌的诊断性DNA探针。
