应用抑制削减杂交技术进行鼠疫耶尔森菌的比较基因组研究

A suppression subtractive hybridization analysis of genomic differences among strains of Yersinia pestis

目的 确定古典型鼠疫耶尔森菌的特有序列 ,为建立和完善该菌基因组分型系统提供可靠数据。方法 通过抑制削减杂交技术发现差异片段并应用PCR验证。结果 发现了 5个在不同生物型鼠疫菌间存在差异的DNA片段 ,其中一个 383bp的片段在来自天山山地灰旱獭、长尾黄鼠鼠疫自然疫源地的 30株鼠疫菌全部阳性 ,而来自其它鼠疫自然疫源地的菌株全部阴性 ,5株假结核耶尔森菌中有 2株 (生物Ⅰ型和Ⅱ型 )阳性。结论 该 383bp片段为天山山地灰旱獭、长尾黄鼠鼠疫自然疫源地的鼠疫菌所特有 ,此疫源地的菌株可能是我国较古老的鼠疫菌。

Objective To identify the specific sequences of biovar antiqua for establishing and improving the genomotyping system of Y.pestis . Methods Different regions (DFR) were identified based by suppression subtractive hybridization and then these DFR were confirmed by PCR. Results Five DFRs were found. One of the DFR of 383 bp was specific to biovar antiqua. The 383 bp fragment was only found in strains of Y.pestis isolated from Marmota baibacina Spermophilus undulates Plague Focus of the Tianshan Mountains, but not found in strains from other Plague Foci. It was also found in two (belonging to biotypeⅠ and Ⅱ) of five strains of Y.pseudotuberculosis . Conclusion The 383 bp fragment is specific to strains of Y.pestis from Marmota baibacina Spermophilus undulates Plague Focus of the Tianshan Mountains and these strains may be the ancestor of Y.pestis in China.