Difference between revisions of "Part:BBa K3606006"
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<partinfo>BBa_K3606006 SequenceAndFeatures</partinfo> | <partinfo>BBa_K3606006 SequenceAndFeatures</partinfo> | ||
<h2>Results:</h2> | <h2>Results:</h2> | ||
| − | https://2020.igem.org/ | + | https://2020.igem.org/wiki/images/4/46/T--Fudan--aTc_400.jpg |
Under the induction of aTc, the expression level of ptetO2 is very low. | Under the induction of aTc, the expression level of ptetO2 is very low. | ||
| − | https://2020.igem.org/ | + | https://2020.igem.org/wiki/images/0/0f/T--Fudan--rate_400.jpg |
| − | + | ||
We have measured the expression level of PtetO2, PtetO3([[BBa_K3606006]]) along with the original PtetR([[BBa_R0040]]) via plate reader by combining them with GFP, when they were induced by different concentration of hydrotetracycline or not induced. | We have measured the expression level of PtetO2, PtetO3([[BBa_K3606006]]) along with the original PtetR([[BBa_R0040]]) via plate reader by combining them with GFP, when they were induced by different concentration of hydrotetracycline or not induced. | ||
| − | + | ||
<h2>Further Application:</h2> | <h2>Further Application:</h2> | ||
Revision as of 18:09, 27 October 2020
PtetO2 promoter
This part is a fusion promoter of PL and tetO, regulated by tetR and anhydrotetracycline
Background:
As a well-characterized promoter, PtetR has been studied thoroughly with its expression level and leakage. To better optimize the antimicrobial peptide expressing system, here we take a unique idea to created a new promoter fusion to create a new promoter that are regulated by tetracycline while with a desired expression intensity.
Design:
This part is a fusion of tetO operon and the -35 upstream, -35--10 parts of the stronger PL promoters in lambda phages, to create a improved promoter with proper expression level while regulated by tetracycline.
Usage and Biology:
This part functions to lower the leakage and adjust the level of expression of the downstream gene. We fuse the -35 upstream, -35--10 parts of the stronger PL promoters in lambda phages with the tetO operon regulated by tetracycline. As a promoter fusion of PL and tetO, not only is it regulated by tetR and anhydrotetracycline, but it also inherited the expression level of PL.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Results:
Under the induction of aTc, the expression level of ptetO2 is very low.
We have measured the expression level of PtetO2, PtetO3(BBa_K3606006) along with the original PtetR(BBa_R0040) via plate reader by combining them with GFP, when they were induced by different concentration of hydrotetracycline or not induced.
Further Application:
This part will act as a tighter tetracycline-regulated promoter with _____ expression level than the original PtetR. The method that applied here can also change the future train of thoughts to create new fusion promoter with different characteristic.
References:
Huanna Qiu, et al. Construction of promoters with tight regulation on chromosome of Escherichia coli. Microbiology China,2018,45(08):1693-1704. (Chinese)