Construction of 2,4,6-Trinitrotoluene Biosensors with Novel Sensing Elements from Escherichia coli K-12 MG1655

被引:0
|
作者
Junjie Tan
Naipeng Kan
Wei Wang
Jingyi Ling
Guolong Qu
Jing Jin
Yu Shao
Gang Liu
Huipeng Chen
机构
[1] Beijing Institute of Biotechnology,College of Life Sciences
[2] Anhui University,College of Life Sciences
[3] Jilin University,undefined
[4] ShenYang Pharmaceutical University,undefined
来源
Cell Biochemistry and Biophysics | 2015年 / 72卷
关键词
Synthetic biology; Biosensor; TNT; GFP; Promoter;
D O I
暂无
中图分类号
学科分类号
摘要
Detection of 2,4,6-trinitrotoluene (TNT) has been extensively studied since it is a common explosive filling for landmines, posing significant threats to the environment and human safety. The rapid advances in synthetic biology give new hope to detect such toxic and hazardous compounds in a more sensitive and safe way. Biosensor construction anticipates finding sensing elements able to detect TNT. As TNT can induce some physiological responses in E. coli, it may be useful to define the sensing elements from E. coli to detect TNT. An E. coli MG1655 genomic promoter library containing nearly 5,400 elements was constructed. Five elements, yadG, yqgC, aspC, recE, and topA, displayed high sensing specificity to TNT and its indicator compounds 1,3-DNB and 2,4-DNT. Based on this, a whole cell biosensor was constructed using E. coli, in which green fluorescent protein was positioned downstream of the five sensing elements via genetic fusion. The threshold value, detection time, EC200 value, and other aspects of five sensing elements were determined and the minimum responding concentration to TNT was 4.75 mg/L. According to the synthetic biology, the five sensing elements enriched the reservoir of TNT-sensing elements, and provided a more applicable toolkit to be applied in genetic routes and live systems of biosensors in future.
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页码:417 / 428
页数:11
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