Difference between revisions of "Part:BBa K3447101"
 
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<partinfo>BBa_K3447133 short</partinfo>
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<partinfo>BBa_K3447101 short</partinfo>
  
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This part can turn off the expression of the sfGFP gene when blue light irradiation.
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RelE is from <i>Escherichia coli</i> str. K-12, encoding RelE toxin.
  
  
 
===Usage and Biology===
 
===Usage and Biology===
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YF1 is the kinase for FixJ in the blue light system. Without blue light irradiation, YF1 phosphorylates FixJ, activating the downstream expression after promoter P<sub>FixK2</sub>. Once the blue light is on, the FixJ cannot be phosphorylated, shutting down the downstream gene expression.<br>
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In our project, we use this part to achieve the response of blue light and inhibit the growth of bacteria.<br>
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===Characterization===
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[[Image: Pathway of Blue Light-on System.jpg|thumb|center|500px|<b>Fig. 1 Pathway of Blue Light-on System.</b> (A) without blue light; (B) with blue light.]]<br>
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Arabinose induced RelE expression: <partinfo>BBa_K3447104</partinfo><br>
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To enrich the blue-light sensing system, repressor pair CI-P<sub>lambda</sub> were introduced in our project to achieve the goal of blue light-on switch system (click to [https://2020.igem.org/Team:Jilin_China/Improvement Jilin_China 2020 Improvement]). Therefore, in our blue light-on system, when blue light is on, P<sub>FixK2</sub> is blocked and P<sub>lambda</sub> can turn on the expression of sfGFP (Fig. 1A). Without blue light, CI functions normally thereby inhibiting the expression of downstream sfGFP (Fig. 1B).<br>
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Based on the experiments above, to verify the function of blue light-on system, two control groups with mock were set to proof the activation of the blue light on our bacteria. As is shown in Fig. 2, compared with the construct without blue light, our blue light-off system would constantly activate the fluorescent expression with blue light induced.
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AHL induced RelE expression: <partinfo>BBa_K3447106</partinfo><br>
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[[Image: Blue Light-on system can regulate the downstream gene via switch of light.jpg|thumb|center|500px|<b>Fig. 2 Blue Light-on system can regulate the downstream gene via switch of light.</b> (A) The emission intensity at 528 nm was measured at the excitation wavelength of 485 nm. After that, measure these values at the indicated time. (B) <i>E. coli</i> DH5α with blue light-on system. (a) 18-hour incubation without blue light; (b) 18-hour incubation with blue light.]]<br>
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==<b>Design</b>==
 
==<b>Design</b>==
 
===Design Notes===
 
===Design Notes===
 
We added some synonymous mutations to avoid part rules.<br>
 
We added some synonymous mutations to avoid part rules.<br>
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K3447133 SequenceAndFeatures</partinfo>
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<partinfo>BBa_K3447101 SequenceAndFeatures</partinfo>
  
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  
 
===Functional Parameters===
 
===Functional Parameters===
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<partinfo>BBa_K3447133 parameters</partinfo>
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<partinfo>BBa_K3447101 parameters</partinfo>
 
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Latest revision as of 10:05, 27 October 2020


RelE, a kind of toxin protein

RelE is from Escherichia coli str. K-12, encoding RelE toxin.


Usage and Biology

In our project, we use this part to achieve the response of blue light and inhibit the growth of bacteria.

Arabinose induced RelE expression: BBa_K3447104

AHL induced RelE expression: BBa_K3447106

Design

Design Notes

We added some synonymous mutations to avoid part rules.



Source

We found this sequence data in GenBank.

References

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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1
    Illegal BsaI.rc site found at 467