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Monitoring arsenic using genetically encoded biosensors in vitro: The role of evolved regulatory genes

文献类型：期刊论文

作者Wang, Xuanyu1,4,5; Zhu, Kaili1,4,5; Chen, Dongdong2; Wang, Juan1,4,5; Wang, Xiaofei3; Xu, An1,4,5; Wu, Lijun1,4,5; Li, Luzhi3; Chen, Shaopeng1,5
刊名ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY
出版日期2021
卷号207
关键词Toxic pollutant Genetically encoded biosensors In vitro Evolved regulatory genes Arsenic
ISSN号0147-6513
DOI10.1016/j.ecoenv.2020.111273
通讯作者Li, Luzhi() ; Chen, Shaopeng(spchen035@ipp.ac.cn)
英文摘要Toxic pollutant (TP) detection in situ using analytical instruments or whole-cell biosensors is inconvenient. Designing and developing genetically coded biosensors in vitro for real-world TP detection is a promising alternative. However, because the bioactivity and stability of some key biomolecules are weakened in vitro, the response and regulation of reporter protein become difficult. Here, we established a genetically encoded biosensor in vitro with an arsenical resistance operon repressor (ArsR) and GFP reporter gene. Given that the wildtype ArsR did not respond to arsenic and activate GFP expression in vitro, we found, after screening, an evolved ArsR mutant ep3 could respond to arsenic and exhibited an approximately 3.4-fold fluorescence in-crease. Arsenic induced expression of both wildtype ArsR and ep3 mutant in vitro, however, only ep3 mutant regulated the expression of reporter gene. Furthermore, the effects of cell extracts, temperature, pH, incubation, and equilibrium time were investigated, and the equilibration of reaction mixtures for 30 min at 37 degrees C was found to be essential for in vitro arsenic detection prior to treatment with arsenic. Based on our data, we established a standard procedure for arsenic detection in vitro. Our results will facilitate the practical application of genetically encoded biosensors in TP monitoring.
WOS关键词FREE PROTEIN-SYNTHESIS ; CELL-FREE BIOSENSOR ; SYNTHETIC BIOLOGY ; SYSTEM
资助项目National Natural Science Foundation of China[21707144] ; Anhui Provincial Natural Science Foundation[1808085QB49] ; Anhui Provincial Natural Science Foundation[1808085QB37] ; Anhui Provincial Key RD Programmes[202004i07020016] ; Key Program of 13th five-year plan, CASHIPS[KP-2017-05] ; Opening Foundation of Anhui Province of Key Laboratory of Environmental Toxicology and Pollution Control Technology[Y79AH72588]
WOS研究方向Environmental Sciences & Ecology ; Toxicology
语种英语
WOS记录号WOS:000592454600004
出版者ACADEMIC PRESS INC ELSEVIER SCIENCE
资助机构National Natural Science Foundation of China ; Anhui Provincial Natural Science Foundation ; Anhui Provincial Key RD Programmes ; Key Program of 13th five-year plan, CASHIPS ; Opening Foundation of Anhui Province of Key Laboratory of Environmental Toxicology and Pollution Control Technology
源URL[http://ir.hfcas.ac.cn:8080/handle/334002/105439]  
专题中国科学院合肥物质科学研究院
通讯作者Li, Luzhi; Chen, Shaopeng
作者单位1.Key Lab Environm Toxicol & Pollut Control Technol, Hefei 230031, Anhui, Peoples R China
2.Anhui Univ, Inst Phys Sci & Informat Technol, Hefei 230601, Anhui, Peoples R China
3.Hefei Univ, Sch Biol Food & Environm, Hefei, Anhui, Peoples R China
4.Univ Sci & Technol China, Hefei 230026, Anhui, Peoples R China
5.Chinese Acad Sci, Hefei Inst Phys Sci, Key Lab High Magnet Field & Ion Beam Phys Biol, Hefei 230031, Anhui, Peoples R China
推荐引用方式
GB/T 7714		 Wang, Xuanyu,Zhu, Kaili,Chen, Dongdong,et al. Monitoring arsenic using genetically encoded biosensors in vitro: The role of evolved regulatory genes[J]. ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY,2021,207.
APA Wang, Xuanyu.,Zhu, Kaili.,Chen, Dongdong.,Wang, Juan.,Wang, Xiaofei.,...&Chen, Shaopeng.(2021).Monitoring arsenic using genetically encoded biosensors in vitro: The role of evolved regulatory genes.ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY,207.
MLA Wang, Xuanyu,et al."Monitoring arsenic using genetically encoded biosensors in vitro: The role of evolved regulatory genes".ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY 207(2021).

入库方式： OAI收割

来源：合肥物质科学研究院

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