Abstract
Small molecule metabolites and their allosterically regulated repressors play an important role in many gene expression and metabolic disorder processes. These natural sensors, though valuable as good logic switches, have rarely been employed without transcription machinery in cells. Here, two pairs of repressors, which function in opposite ways, were cloned, purified and used to control DNA replication in rolling circle amplification (RCA) in vitro. By using metabolites and repressors as inputs, RCA signals as outputs, four basic logic modules were constructed successfully. To achieve various logic computations based on these basic modules, we designed series and parallel strategies of circular templates, which can further assemble these repressor modules in an RCA platform to realize twelve two-input Boolean logic gates and a three-input logic gate. The RCA-output and RCA-assembled platform was proved to be easy and flexible for complex logic processes and might have application potential in molecular computing and synthetic biology.
Highlights
S-adenosylmethionine (SAM) in lung cancer[20]
Rolling circle amplification (RCA) has derived a variety of signal output means, such as fluorescence, colorimetric, electronic, physicochemical and nanopore analyses[27,28,29,30,31], which are extremely desirable in logic operations
When LacI was added, it bound the circular template by the recognition sequence, and prohibited DNA polymerase from RCA
Summary
S-adenosylmethionine (SAM) in lung cancer[20]. In these cases, metabolite-regulated allosteric TFs might provide another alternative route for metabolites assays. When LacI was added, it bound the circular template by the recognition sequence, and prohibited DNA polymerase from RCA. When IPTG was added, it induced LacI allosterically to release the circular template; the RCA occurred again.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Disclaimer: 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.