Difference between revisions of "Part:BBa K3408010"
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<partinfo>BBa_K3408010 short</partinfo>
 
<partinfo>BBa_K3408010 short</partinfo>
  
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We used promoter P<sub>nar</sub> which was activated by FNR to trigger the expression of GFP. If there was no oxygen, then FNR could be transcribed and we could see green bacteria in our culture medium.
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We designed this composite part by combining promoter P<sub>nar</sub>, trigger RNA, P<sub>CⅠ</sub>, switch RNA and mazF. So we could utilize it to verify the feasibility of our device, which comprises of Toehold switch and suicide protein—mazF. Former composite part BBa_K3408006 has demonstrated the feasibility of promoter P<sub>nar</sub>, so when our engineered bacteria <i>Bacillus subtilis</i> is in anaerobic environment, promoter P<sub>nar</sub> is activated by global regulator—FNR. Then downstream trigger RNA is expressed. As switch RNA is constantly expressed under the control of promoter P<sub>C1</sub>, the trigger RNA can successfully trigger the Toehold switch, thus leading to the expression of mazF. So <i>Bacillus subtilis</i> can commit suicide. While in aerobic environment, promoter P<sub>nar</sub> ceases to work, Toehold switch cannot be triggered. As a result, mazF can not express. This device proves that our suicide module and Toehold switch can successfully combine together.  
  
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Plasmid profile
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https://2020.igem.org/wiki/images/d/d2/T--NAU-CHINA--composite-parts5.1.png
  
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Plasmid profile
 
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https://2020.igem.org/wiki/images/1/18/T--NAU-CHINA--composite-parts1.1.png
 
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===

Revision as of 14:11, 16 October 2020

Pnar-B0034-Trigger RNA-B0015-PCⅠ-Switch RNA-mazF-B0015

We designed this composite part by combining promoter Pnar, trigger RNA, PCⅠ, switch RNA and mazF. So we could utilize it to verify the feasibility of our device, which comprises of Toehold switch and suicide protein—mazF. Former composite part BBa_K3408006 has demonstrated the feasibility of promoter Pnar, so when our engineered bacteria Bacillus subtilis is in anaerobic environment, promoter Pnar is activated by global regulator—FNR. Then downstream trigger RNA is expressed. As switch RNA is constantly expressed under the control of promoter PC1, the trigger RNA can successfully trigger the Toehold switch, thus leading to the expression of mazF. So Bacillus subtilis can commit suicide. While in aerobic environment, promoter Pnar ceases to work, Toehold switch cannot be triggered. As a result, mazF can not express. This device proves that our suicide module and Toehold switch can successfully combine together.

Plasmid profile T--NAU-CHINA--composite-parts5.1.png

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 187
    Illegal BglII site found at 409
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]