【摘要】 目的在大肠杆菌基因组中筛选可用于TNT检测的基因传感元件。方法利用随机酶切方法不完全酶切大肠杆菌K-12 MG1655基因组,构建基因组启动子文库,通过多轮筛选法获得可感应TNT的文库元件。通过数据库分析,并在灵敏度、特异性、起效时间各方面对新的传感元件在TNT诱导下的启动活性进行考察,进一步确证了文库元件中发挥作用的功能序列。结果和结论成功构建了大肠杆菌K-12 MG1655基因组启动子文库,并从中筛选得到了一个可感应TNT的文库元件,经过进一步分析确证了其发挥功能的序列top Ap4,并在灵敏度、特异性、起效时间上都表现出较好的启动活性。上述研究工作发现了top Ap4对TNT的感应作用,并验证了其具有较好的启动活性,为TNT检测生物传感器的构建打下了良好的基础。更多还原

【Abstract】 Objective To screen the sensing elements for TNT detection in Escherichia coli genome. Methods A genome promoter library with cutting E. coli K-12 MG1655 genome was constructed. Bacterial luciferase lux CDABE was used as a reporter gene during promoter screening. We discovered TNT sensing elements through several rounds of screening. Through analysis of sensitivity,specificity and timeliness,the promoter activity of the elements was evaluated,and the functional sequence of the elements was further confirmed. Results and Conclusion We successfully constructed an E. coli K-12 MG1655 genome library,from which a TNT sensing element was discovered,which had a good performance in the analysis of sensitivity,specificity and timeliness. In this study,we reported that the top Ap4 is a TNT sensing element for the first time. We also verified its excellent promoter activity.更多还原