Bioluminescence detection system of mutagen using firefly luciferase genes introduced in Escherichia coli lysogenic strain
Bioluminescence detection system of mutagen using firefly luciferase genes introduced in Escherichia coli lysogenic strain
复制标题
利用大肠杆菌溶原菌株中引入的萤火虫荧光素酶基因诱变剂生物发光检测系统
DOI：
10.1021/ac00041a004
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发表时间：
1992
影响因子：
7.4
通讯作者：
I. Karube
中科院分区：
文献类型：
--
作者：
Soo Mi Lee;Masayasu Suzuki;M. Kumagai;H. Ikeda;E. Tamiya;I. Karube
利用大肠杆菌溶源性菌株的原噬菌体诱导和生物发光技术，建立了一种快速、简便的诱变剂微生物传感系统。该系统由溶原性E.编码萤火虫荧光素酶基因的大肠杆菌和光检测系统。诱变剂丝裂霉素C的测定是通过测定E.大肠杆菌溶原性菌株中的发光底物的存在下的重组噬菌体。在加入丝裂霉素C后约1小时，开始观察到发光，3小时后，其发光水平达到1小时的2倍。紫外线照射也产生光的基础上诱导噬菌体从大肠杆菌。大肠杆菌溶原性菌株为重组噬菌体。另一方面，当向反应介质中加入非致突变毒性化合物（如叠氮化钠）时，未观察到发光。丝裂霉素C可以在1小时内检测到这种传感系统，在浓度低至10（2）ng/测定。
A rapid and convenient microbial sensing system for mutagens was developed based upon the induction of prophage from Escherichia coli lysogenic strain and bioluminescence. The system consisted of lysogenic E. coli encoding firefly luciferase genes and a photodetection system. Measurement of mutagen mitomycin C was achieved by measuring the luminescence intensity emitted from E. coli lysogenic strain for the recombinant phage in the presence of luminescence substrates. Approximately 1 h after addition of mitomycin C, the luminescence began to be observed, and 3 h after, it attained a level of 2 times greater than that of 1 h. Irradiation with ultraviolet light also produced light based on induction of phage from the E. coli lysogenic strain for the recombinant phage. On the other hand, when nonmutagenic toxic compounds like sodium azide were added to the reaction medium, luminescence was not observed. Mitomycin C could be detected within 1 h with this sensing system, at concentrations down to 10(2) ng/assay.