Difference between revisions of "Part:BBa K3606006"
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This part is a fusion promoter of PL and tetO, regulated by tetR and anhydrotetracycline
 
This part is a fusion promoter of PL and tetO, regulated by tetR and anhydrotetracycline
 +
<h2>Background:</h2>
 +
To better establish the antimicrobial system in our design, we
  
 
<h2>Design:</h2>
 
<h2>Design:</h2>
−
This part functions to lower the leakage and raise the level of expression of the downstream gene. As a well-characterized promoter, ptetR has been studied thoroughly with its expression level and leakage. While here we take a unique idea to created a new promoter fusion: 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.  
+
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.
 +
 
 +
<h2>Usage and Biology:</h2>
 +
This part functions to lower the leakage and adjust the level of expression of the downstream gene.As a well-characterized promoter, ptetR has been studied thoroughly with its expression level and leakage. While here we take a unique idea to created a new promoter fusion: 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.  
  
−
<!-- Add more about the biology of this part here
 
−
===Usage and Biology===
 
  
 
<!-- -->
 
<!-- -->
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K3606006 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3606006 SequenceAndFeatures</partinfo>
 +
<h2>Results:</h2>
 +
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
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<h2>Further Application:</h2>
 +
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.
  
 +
<h2>References:</h2>
 +
Yingna Qiu, et al. The construction of rigorous type promoter,i on E. coli chromosomes. Microbiology China,2018,45(08):1693-1704.
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  

Revision as of 08:26, 25 October 2020


PtetO2 promoter

This part is a fusion promoter of PL and tetO, regulated by tetR and anhydrotetracycline

Background:

To better establish the antimicrobial system in our design, we

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.As a well-characterized promoter, ptetR has been studied thoroughly with its expression level and leakage. While here we take a unique idea to created a new promoter fusion: 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_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.