Abstract
Antibiotics as emerging environmental contaminants, are widely used in both human and veterinary medicines. A solid-state nanopore sensing method is reported in this article to detect Tetracycline, which is based on Tet-off and Tet-on systems. rtTA (reverse tetracycline-controlled trans-activator) and TRE (Tetracycline Responsive Element) could bind each other under the action of Tetracycline to form one complex. When the complex passes through nanopores with 8 ~ 9 nanometers in diameter, we could detect the concentrations of Tet from 2 ng/mL to 2000 ng/mL. According to the Logistic model, we could define three growth zones of Tetracycline for rtTA and TRE. The slow growth zone is 0–39.5 ng/mL. The rapid growth zone is 39.5−529.7 ng/mL. The saturated zone is > 529.7 ng/mL. Compared to the previous methods, the nanopore sensor could detect and quantify these different kinds of molecule at the single-molecule level.
Highlights
Tet-Off and Tet-On systems are inducible expression models for studies on eukaryote cells and organisms[9,10,11]
DNA binding with reverse tet-transactivator (rtTA) protein in present of Tet with the Electrophoretic Mobility Shift Assay (EMSA) method
The target DNA of tetracycline-responsive element (TRE) fragment is ~300 bp oligonucleotide duplex containing a minicytomegalovirus (CMV) promoter and TRE full-length sequence that could bind to rtTA with Tet in vivo
Summary
Tet-Off and Tet-On systems are inducible expression models for studies on eukaryote cells and organisms[9,10,11]. TTA can induce gene expression from TRE promoters in eukaryotic cells, while in the presence of Tet or dox, that inactivates the tTA-TRE interaction and switches gene expression off (Tet-Off system)[12] In another Tet system (Tet-On system), rtTA can only recognize TRE-regulated target gene to promote gene expression with Tet or dox-dependent[13]. It has been two decades since the beginning from the study of nanopore-based single-molecule sensor, which could sense protein or DNA molecule by a channel connected by two chambers filled with electrolyte solution[14,15]. We could identify the presence of tetracycline with solid-state nanopore technique
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
