Flowchart - Adaptor characterised with a RNA Thermometer
The principle behind the adaptor is based on the fact that, translation of the leader peptide in the tnaC element determines whether or not the genes further downstream get transcribed(Konan et al,2000). The tnaC element in the tna operon is followed by a Rho binding site, and when translation occurs the rho factor does not bind to the rho binding site leading to transcription antitermination(Transcription occurs depending on whether or not translation of tnaC has occured) . Hence by replacing the constitutive RBS upstream of the tnaC element(tnaC pepetide + Rho factor binding sites) with a translational regulatory riboswitch of our choice we can convert the translational regulation into transcriptional regulation (Chang.C.Liu et al,2012). To test the adaptor we will place the before mentioned RNA thermometer and pH riboswitch, upstream of the adaptor and GFP reporter protein downstream of the adaptor.
To test the functioning of the adaptor we will place the above mentioned RNA thermometer and pH riboswitch upstream of the leader peptide element and a reporter protein downstream of the leader peptide element. This adaptor can further be used by any team which intends to convert the translational regulatory property of a riboswitch to transcriptional regulation. Furthermore, this adaptor is both easy to engineer and can be easily assembled into higher order function.
Characterization studies can be found here.
Adaptor with RNA pH Meter
Adaptor with RNA Thermometer
- Konan, K.V. & Yanofsky, C. Rho-Dependent transcription termination in the tna Operon of Escherichia coli: Roles of the boxA Sequence and the rut Site.J. Bacteriol. 182(14), 3981–3988 (2000).
- Chang, C. L.,Lei Qi, Lucks J.B., Segall-Shapiro T.H., Wang, D.,Mutalik V.K.& Arkin, A.P.An adaptor from translational to transcriptional control enables predictable assembly of complex regulation. Nat. Methods 9,1088–1094(2012).
- Richardson J.P. Loading Rho to terminate transcription. Cell 114, 157–159 (2003).