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Part:BBa_K3606006
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:
We have measured the expression level of PtetO2, PtetO3(BBa_K3606007) along with the original PtetR(BBa_R0040)when they were induced by different concentration of hydrotetracycline or not induced. Both of these promoters exhibit good low leakage characteristics. Compared to ptetR, their peak intensity can also reach a more proper expression level for our system. 图
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:
Yingna Qiu, et al. The construction of rigorous type promoter,i on E. coli chromosomes. Microbiology China,2018,45(08):1693-1704.
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