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OALib Journal期刊
ISSN: 2333-9721
费用：99美元
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缺失hpi毒力岛的eaggeco42突变菌株的构建及鉴定
, PP. 1352-1356
Keywords: 肠集聚性粘附大肠杆菌,hpi毒力岛,耶尔森菌,缺失
Abstract:
目的构建并鉴定hpi毒力岛缺失的eaggec突变株。方法运用基因重组方法,以irp8基因部分序列作为同源重组的一侧序列,irp5基因序列作为同源重组的另一侧序列,中间插入有卡那霉素(km)抗性基因(kan)标记。以pcvd442作为载体,构建含有irp8部分序列和irp5基因的重组自杀质粒pco85。以eaggeco42为出发菌株,构建缺失irp8～irp5约24kb的hpi毒力岛功能核心区区域的全岛缺失株eag85。结果通过接合转移和同源重组,利用蔗糖抗性筛选,pco85上的同源序列有效的置换了eaggeco42的irp8和irp5基因。pcr方法扩增irp3、ybta、irp9等基因的结果显示,筛选出的eag85确为缺失hpi毒力岛基因的eaggec菌株。结论成功地构建了缺失hpi毒力岛的eaggeco42突变菌株,为进一步阐明hpi毒力岛在eaggec菌株中的功能建立了重要的物质基础。
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