Difference between revisions of "Part:BBa K3202068"
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In our system(Fig. 1), we have transcriptional repressors R1 and R2 expressed through the inducible promoter but translationally inhibited by the MicC sRNA1 and sRNA2 respectively, since RBS2 and RBS3 is directly inhibited by sRNA while they initiates the transcription of the two repressors; the gene of interest is translationally coupled to the repressor gene while three promoters which produces the inhibitory sRNA are targeted by their corresponding repressors. Therefore, repressors and sRNAs are mutually inhibitory. | In our system(Fig. 1), we have transcriptional repressors R1 and R2 expressed through the inducible promoter but translationally inhibited by the MicC sRNA1 and sRNA2 respectively, since RBS2 and RBS3 is directly inhibited by sRNA while they initiates the transcription of the two repressors; the gene of interest is translationally coupled to the repressor gene while three promoters which produces the inhibitory sRNA are targeted by their corresponding repressors. Therefore, repressors and sRNAs are mutually inhibitory. | ||
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Revision as of 10:18, 20 October 2019
AraC-Pc-pBAD-RBS1-Colicin E2-T500-T1/TE-MicCsRNA-PA1/O4
This composite part is designed to insert the toxin Colicin E2 into the bistable system.
Presenting with BBa_K3202066-BBa_K3202067, this part is to test the function of the toxin within our bistable system.
Background
We are inspired by a lately published article written by Belén Calles, Angel Goñi-Moreno, and Víctor de Lorenzo to design a double-bistable system which achieves zero basal expression of gene of interest in the absence of any inducer while ensuring constant transcriptional capacities and induction rates among promoters.
“The mechanism of a bistable system is elaborated below: a repressor transcriptionally inhibits the promoter which is responsible for the transcription of sRNA, while the sRNA translationally inhibits the repressor. The gene of interest is translationally coupled to the repressor gene. That is to say, the repressor protein R targets the strong promoter which the inhibitory sRNA is produced, so that R and sRNA are mutually inhibitory. Such arrangement helps to minimize the basal activity of the inducible module and thus suppress leaky gene expression levels.”
Design
In our system(Fig. 1), we have transcriptional repressors R1 and R2 expressed through the inducible promoter but translationally inhibited by the MicC sRNA1 and sRNA2 respectively, since RBS2 and RBS3 is directly inhibited by sRNA while they initiates the transcription of the two repressors; the gene of interest is translationally coupled to the repressor gene while three promoters which produces the inhibitory sRNA are targeted by their corresponding repressors. Therefore, repressors and sRNAs are mutually inhibitory.
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Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1144
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 979
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 961