By virtue of genetic engineering technology,we developed a highly sensitive biosensor for tetracycline detection based on the interaction between the tetracycline regulator TetR and tetracycline.In the absence of tetr...By virtue of genetic engineering technology,we developed a highly sensitive biosensor for tetracycline detection based on the interaction between the tetracycline regulator TetR and tetracycline.In the absence of tetracycline,TetR binds to the tetO sequence,inhibiting the expression of the sfGFP reporter gene.When tetracycline is present,it induces TetR to release from tetO,allowing sfGFP expression.The biosensor was optimized through the selection of transcription factors and reporter genes,and the optimization of spacer lengths.Chassis cells were grown to mid-log phase in a tetracycline-supplemented medium for subsequent fluorescence intensity measurement.The biosensor exhibited a strong linear correlation between fluorescence intensity and tetracycline concentration(I=37,620.7×[C(Tc)]+4048.5,R^(2)=0.998),demonstrating high sensitivity with a detection limit of 0.0097 mg/L.The response time of the biosensor ranged from 2 to 4 h within the working concentration range,making it suitable for real-time detection.It shows potential for application in actual water sample analysis and as an early warning technology for water pollution risks.展开更多
生物传感器已经成为实时监测特定小分子和精确控制生物系统中基因表达的强大工具。用于1,4-丁二胺生物合成的高通量传感器可以极大地提高1,4-丁二胺高产菌株的筛选效率。为研究调整生物传感器特性的策略,本研究开发了一种以转录调节因子...生物传感器已经成为实时监测特定小分子和精确控制生物系统中基因表达的强大工具。用于1,4-丁二胺生物合成的高通量传感器可以极大地提高1,4-丁二胺高产菌株的筛选效率。为研究调整生物传感器特性的策略,本研究开发了一种以转录调节因子PuuR为基础的1,4-丁二胺生物传感器,其同源的操作子puuO被组装在大肠杆菌组成型启动子PgapA中,以控制下游的高能绿色荧光蛋白(superfolder green fluorescent protein,sfGFP)作为报告蛋白表达。最终该传感器在1,4-丁二胺浓度处于0–50 mmol/L时GFP/OD600值与1,4-丁二胺浓度之间能稳定地表现出线性关系。本研究采用大肠杆菌基因组中不同强度的启动子对1,4-丁二胺生物传感器进行分子改造,探究并改进基于PuuR的1,4-丁二胺生物传感器的功能性质,为高通量筛选高产1,4-丁二胺的工程菌株奠定了基础。展开更多
Wearable sensors have revolutionized health monitoring by transitioning from clinical diagnostics to continuous,real-time applications in daily life.The oral cavity,rich in saliva containing over 1,000 biomarkers that...Wearable sensors have revolutionized health monitoring by transitioning from clinical diagnostics to continuous,real-time applications in daily life.The oral cavity,rich in saliva containing over 1,000 biomarkers that reflect systemic health(e.g.,glucose,cortisol,and inflammatory markers)[1],offers the advantage of non-invasive sampling.Its superior environmental stability and strong connection to key physiological processes make it an ideal candidate in the field of digital medicine,serving as a natural gateway to personalized health monitoring.Therefore,the oral cavity represents not only a convenient sampling site but also a strategic interface for realizing the vision of continuous,personalized digital health monitoring.展开更多
Glucose,ascorbic acid(AA),uric acid(UA),and dopamine(DA)are vital biomarkers whose dynamic concentrations correlate with critical diseases;however,multiplexed detection remains challenging for conventional electrochem...Glucose,ascorbic acid(AA),uric acid(UA),and dopamine(DA)are vital biomarkers whose dynamic concentrations correlate with critical diseases;however,multiplexed detection remains challenging for conventional electrochemical sensors because of their limited sensitivity and selectivity.Here,we present a millimeter-scale all-poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate)(PEDOT:PSS)organic electrochemical transistor(OECT)platform that integrates dual-mode sensing with enzyme/metal-free operation for ultrasensitive biomarker monitoring.By engineering polycrystalline PEDOT:PSS channels via H_2 SO_4 post-treatment,the device achieves record-high conductivity(about(2312.0±29.9)S·cm^(–1)),maximum transconductance(about(2.82±0.12)mS),and on/off ratio(about 210.0±7.8),enabling signal amplification at low gate voltages.The dual-mode strategy combines the selectivity of electrochemistry with the sensitivity of OECTs,realizing simultaneous detection of glucose,AA,UA,and DA with clinical-level sensitivity:detection limits down to 8 nmol·L^(–1)(glucose),0.5 nmol·L^(–1)(AA),5 nmol·L^(–1)(DA),and 0.5 nmol·L^(–1)(UA).Validation using human urine samples yielded recovery rates of 94%–114%.This flexible sensing platform provides a new pathway for the development of wearable biosensors for precision diagnostics.展开更多
基金supported by Chinese National Key Programs for Fun-damental Research and Development(No.2023YFC3709005).
文摘By virtue of genetic engineering technology,we developed a highly sensitive biosensor for tetracycline detection based on the interaction between the tetracycline regulator TetR and tetracycline.In the absence of tetracycline,TetR binds to the tetO sequence,inhibiting the expression of the sfGFP reporter gene.When tetracycline is present,it induces TetR to release from tetO,allowing sfGFP expression.The biosensor was optimized through the selection of transcription factors and reporter genes,and the optimization of spacer lengths.Chassis cells were grown to mid-log phase in a tetracycline-supplemented medium for subsequent fluorescence intensity measurement.The biosensor exhibited a strong linear correlation between fluorescence intensity and tetracycline concentration(I=37,620.7×[C(Tc)]+4048.5,R^(2)=0.998),demonstrating high sensitivity with a detection limit of 0.0097 mg/L.The response time of the biosensor ranged from 2 to 4 h within the working concentration range,making it suitable for real-time detection.It shows potential for application in actual water sample analysis and as an early warning technology for water pollution risks.
文摘生物传感器已经成为实时监测特定小分子和精确控制生物系统中基因表达的强大工具。用于1,4-丁二胺生物合成的高通量传感器可以极大地提高1,4-丁二胺高产菌株的筛选效率。为研究调整生物传感器特性的策略,本研究开发了一种以转录调节因子PuuR为基础的1,4-丁二胺生物传感器,其同源的操作子puuO被组装在大肠杆菌组成型启动子PgapA中,以控制下游的高能绿色荧光蛋白(superfolder green fluorescent protein,sfGFP)作为报告蛋白表达。最终该传感器在1,4-丁二胺浓度处于0–50 mmol/L时GFP/OD600值与1,4-丁二胺浓度之间能稳定地表现出线性关系。本研究采用大肠杆菌基因组中不同强度的启动子对1,4-丁二胺生物传感器进行分子改造,探究并改进基于PuuR的1,4-丁二胺生物传感器的功能性质,为高通量筛选高产1,4-丁二胺的工程菌株奠定了基础。
基金support from the Guangdong Basic and Applied Basic Research Foundation(Nos.2021A1515110388,2024A1515011707)Science and Technology Projects in Guangzhou(No.2024A04J5195)Shenzhen Natural Science Foundation(Nos.JCYJ20230807111120043).
文摘Wearable sensors have revolutionized health monitoring by transitioning from clinical diagnostics to continuous,real-time applications in daily life.The oral cavity,rich in saliva containing over 1,000 biomarkers that reflect systemic health(e.g.,glucose,cortisol,and inflammatory markers)[1],offers the advantage of non-invasive sampling.Its superior environmental stability and strong connection to key physiological processes make it an ideal candidate in the field of digital medicine,serving as a natural gateway to personalized health monitoring.Therefore,the oral cavity represents not only a convenient sampling site but also a strategic interface for realizing the vision of continuous,personalized digital health monitoring.
基金financially supported by the National Natural Science Foundation of China(Nos.52272214,52372082,52466013,52373184,and U24A20660)Jiangxi Provincial Natural Science Foundation(Nos.20242BAB26059,20232BAB204032,20252BAC200290,20252BEJ730349,and 20252BAC240326)Doctoral Start-Up Fund of Jiangxi Science&Technology Normal University(No.2024BSQD16)。
文摘Glucose,ascorbic acid(AA),uric acid(UA),and dopamine(DA)are vital biomarkers whose dynamic concentrations correlate with critical diseases;however,multiplexed detection remains challenging for conventional electrochemical sensors because of their limited sensitivity and selectivity.Here,we present a millimeter-scale all-poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate)(PEDOT:PSS)organic electrochemical transistor(OECT)platform that integrates dual-mode sensing with enzyme/metal-free operation for ultrasensitive biomarker monitoring.By engineering polycrystalline PEDOT:PSS channels via H_2 SO_4 post-treatment,the device achieves record-high conductivity(about(2312.0±29.9)S·cm^(–1)),maximum transconductance(about(2.82±0.12)mS),and on/off ratio(about 210.0±7.8),enabling signal amplification at low gate voltages.The dual-mode strategy combines the selectivity of electrochemistry with the sensitivity of OECTs,realizing simultaneous detection of glucose,AA,UA,and DA with clinical-level sensitivity:detection limits down to 8 nmol·L^(–1)(glucose),0.5 nmol·L^(–1)(AA),5 nmol·L^(–1)(DA),and 0.5 nmol·L^(–1)(UA).Validation using human urine samples yielded recovery rates of 94%–114%.This flexible sensing platform provides a new pathway for the development of wearable biosensors for precision diagnostics.
