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>
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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.
 
https://2020.igem.org/wiki/images/4/46/T--Fudan--aTc_400.jpg
 
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/wiki/images/0/0f/T--Fudan--rate_400.jpg
 
https://2020.igem.org/wiki/images/0/0f/T--Fudan--rate_400.jpg
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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.  
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PtetO2 has a higher fold of increase than ptetR. This means that it is the more sensitive to the induction of aTc.
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Revision as of 18:12, 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. T--Fudan--img_pteto2.png

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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Results:

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. T--Fudan--aTc_400.jpg Under the induction of aTc, the expression level of ptetO2 is very low. T--Fudan--rate_400.jpg PtetO2 has a higher fold of increase than ptetR. This means that it is the more sensitive to the induction of aTc.


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)