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目的 将绿色荧光特征引入大肠杆菌E coliO15 7∶H7,用于传统检测方法的改进及目的菌的应用研究。方法 将pGFP质粒转化E coliO15 7∶H7,构建一种工程菌 (E coliO15 7∶H7 pGFP) ,并进行了选择性培养基的筛选。以E coliO15 7∶H7 pGFP对鸡肉与牛奶等食品样品进行接种与回收试验 ,同时设定不同温度模拟食品储存的不同情况。结果 pGFP质粒可以在E coliO15 7∶H7菌株中稳定存在。将E coliO15 7∶H7 pGFP接种食品样品 ,以LBan培养基平板回收发现 :较高温度存放肉类与牛奶 ,其污染的E coliO15 7∶H7菌量可在 12h增加 35 0 0 0~ 2 0 0 0 0 0倍 ,而 4℃存放可以使污染菌量缓慢下降。结论 构建了一种具有紫外灯下发出绿色荧光与氨苄青霉素抗性等特性的重组菌株———O15 7∶H7 pGFP ,并设计了适合E coliO15 7∶H7 pGFP生长的选择性培养基———LBan。该重组菌株可应用于检测方法的改进及该菌特性的研究 ,是一种非常有用的工程菌
Objective To introduce green fluorescence into Escherichia coli E. coli O15 7:H7 for the improvement of traditional detection methods and the application of target bacteria. Methods The pGFP plasmid was transformed into E. coli O15 7:H7 to construct an E. coli O15 7:H7 pGFP and the selective medium was screened. E coliO15 7:H7 pGFP on chicken and milk food samples for vaccination and recovery tests, set different temperatures simulate different conditions of food storage. As a result, the pGFP plasmid was stably present in the E coli O15 7: H7 strain. The E coliO15 7:H7 pGFP inoculated food samples to LBan medium flat recovery found: higher temperature storage of meat and milk, the contamination of the E coliO15 7:H7 bacteria can be increased at 12h 35 0 0 0 ~ 2 0 0 0 0 0 times, and 4 ℃ storage can make the amount of bacteria slowly decreased. Conclusion A recombinant strain O15 7:H7 pGFP with the characteristics of green fluorescence and ampicillin resistance was constructed under ultraviolet light and a selective medium suitable for the growth of E coli O15 7:H7 pGFP was designed. LBan. The recombinant strain can be applied to improve the detection method and study the characteristics of the bacterium, and is a very useful engineering bacterium