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

doi:10.1016/j.ecoenv.2020.111273

	Monitoring arsenic using genetically encoded biosensors in vitro: The role of evolved regulatory genes
	Wang, Xuanyu 1,4,5; Zhu, Kaili 1,4,5; Chen, Dongdong 3; Wang, Juan 1,4,5; Wang, Xiaofei 2; Xu, An 1,4,5; Wu, Lijun 1,4,5; Li, Luzhi 2; Chen, Shaopeng 1,4
刊名	ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY
	2021
卷号	207
关键词	Toxic pollutant Genetically encoded biosensors In vitro Evolved regulatory genes Arsenic
ISSN号	0147-6513
DOI	10.1016/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.
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关键词	FREE PROTEIN-SYNTHESIS ; CELL-FREE BIOSENSOR ; SYNTHETIC BIOLOGY ; SYSTEM
WOS研究方向	Environmental Sciences & Ecology ; Toxicology
语种	英语
出版者	ACADEMIC PRESS INC ELSEVIER SCIENCE
WOS记录号	WOS:000592454600004
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
内容类型	期刊论文
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.Chinese Acad Sci, Hefei Inst Phys Sci, Key Lab High Magnet Field & Ion Beam Phys Biol, Hefei 230031, Anhui, Peoples R China 2.Hefei Univ, Sch Biol Food & Environm, Hefei, Anhui, Peoples R China 3.Anhui Univ, Inst Phys Sci & Informat Technol, Hefei 230601, Anhui, Peoples R China 4.Key Lab Environm Toxicol & Pollut Control Technol, Hefei 230031, Anhui, Peoples R China 5.Univ Sci & Technol China, Hefei 230026, 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).