【作者】 王斌；

【导师】 叶冬青； 王红；

【作者基本信息】 安徽医科大学 ， 流行病与卫生统计学， 2001， 硕士

【摘要】 本次研究，从Genebank中调出小肠结肠炎耶尔森菌WA型（YenWA）HPI毒力岛中的irp1、irp3和irp4基因的序列，用Goldkey软件设计了它们的引物。运用分子生物学的PCR技术与原位点杂交技术，对这三个基因在大肠杆菌中的分布状况进行了检测研究。 这是国内外首次对大肠杆菌中HPI毒力岛的irp1、irp3和irp4基因分布进行的大量检测研究，并对结果予以了描述与分析，期望为HPI毒力岛在耶尔森菌与大肠杆菌间存在水平性转移提供必要的客观实验依据。 两种分子生物学技术检测结果基本相一致：106株大肠杆菌中，发现（1）13株存在irp1基因，阳性率为12.3％；（2）92株存在irp3基因，阳性率为86.8％；（3）65株存在irp4基因，阳性率为61.3%（注：以PCR检测结果为准）。这说明大肠杆菌中存在着小肠结肠炎耶尔森菌HPI毒力岛的irp1、irp3和irp4基因，但三基因在大肠杆菌中的分布率有明显性差异。 作者据此提出以下观点：（1）小肠结肠耶尔森菌和大肠杆菌中国株间存在着HPI毒力岛的irp1、irp3和irp4基因的转移，证实了国内外研究关于这两种菌种之间存在着HPI毒力岛水平性转移的报道；（2）转移过程中，irp1比irp3、irp4基因的缺失率要大得多，irp3和irp4基因的缺失率则较为接近；（3）PCR和核酸分子杂交技术是基因检测可行、敏感、有效的基本分子生物学手段。更多还原

【Abstract】 In this study, we investigate the distributions of irpl, irp3 and irp4 genes located Yersinia enterocolitia HPI by PCR amplification and situ dot hybridization assay. This is the first study on the distributions of irpl, irp3, and irp4 genes of the HPI located on Yersinia enterocolitia among so many E.coli strains.And we describe and analysis the results to provide a necessary experimental base on horizontal transfer of the HPI between Y enterocolitia and some E.coli strains. With goldkey software, we design three pairs primers of irpl, irp3 and ipr4 genes of the Yen HPI from Genebank. And detect the distributions of the three genes among 106 E.coli strains by PCR smplification and situ dot hybridization method respectively. Both the two results detected in this study are identical(the results of PCR): (1) Thirteen strains are positive for irpl gene(12.3%); (2)Ninty-two strains are positive for irp3 gene(86.8%); (3)Sixty-five strains are positive for irp4 gene(6 1.3%). These results demonstrate that the three genes exist among E. ccli strains. Accordingly, the author conclude that:Firstly ,the horizontal tranfers of irpl, irp3, and irp4 genes between Yen and some E.coli strains do exist. Secon dly, the deletion rate of irpl gene is far higher than that of irp3 gene or irp4 gene. Thirdly, both PCR and hybridization technologies are feasible, sensitive and effective to detect genes.更多还原