Difference between revisions of "Part:BBa K3408008"
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<partinfo>BBa_K3408008 short</partinfo>
 
<partinfo>BBa_K3408008 short</partinfo>
  
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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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Used P<sub>nar</sub>(BBa_K3408000), RBS(BBa_B0034), optimized CⅠ(BBa_K3408004), terminator(BBa_B0015), P<sub>CⅠ</sub>(BBa_R0051), RBS(BBa_B0034) and GFP(BBa_E0040) to express GFP when PCⅠ was activated. When our <i>Bacillus subtilis</i> was in an aerobic environment, P<sub>nar</sub> was suppressed without FNR and no CⅠ protein could be expressed so that P<sub>CⅠ</sub> could be activated and we could see GFP expressed. But when our <i>Bacillus subtilis</i> was in an anerobic environment, CⅠ protein could be expressed to suppress P<sub>CⅠ</sub> and we couldn't see GFP anymore.
  
 
Plasmid profile
 
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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https://2020.igem.org/wiki/images/e/e9/T--NAU-CHINA--composite-parts3.1.png
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<!-- 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 13:54, 16 October 2020

Pnar-B0034-CⅠ-B0015-PCⅠ-B0034-GFP-B0015

Used Pnar(BBa_K3408000), RBS(BBa_B0034), optimized CⅠ(BBa_K3408004), terminator(BBa_B0015), PCⅠ(BBa_R0051), RBS(BBa_B0034) and GFP(BBa_E0040) to express GFP when PCⅠ was activated. When our Bacillus subtilis was in an aerobic environment, Pnar was suppressed without FNR and no CⅠ protein could be expressed so that PCⅠ could be activated and we could see GFP expressed. But when our Bacillus subtilis was in an anerobic environment, CⅠ protein could be expressed to suppress PCⅠ and we couldn't see GFP anymore.

Plasmid profile T--NAU-CHINA--composite-parts3.1.png

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.rc site found at 1709