Difference between revisions of "Part:BBa K3408011"

Line 1: Line 1:
 
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K3408011 short</partinfo>
 
<partinfo>BBa_K3408011 short</partinfo>
  
We used constitutive promoter P<sub>liaG</sub> to transcribe trigger RNA all the time. And we also used P<sub>C&#8544;</sub> as a constitutive promoter to transcribe switch RNA. If our Toehold switch could work normally, GFP could be translated and we could see green bacteria in our culture medium.
+
This composite part consists of promoter P<sub>liaG</sub> and P<sub>grac</sub>, lacI gene, CⅠ gene, GFP gene and some essential RBS and terminators. Promoter P<sub>liaG</sub> starts transcription process and its transcription production is LacI, which is regulated by IPTG. When bacteria is exposed to medium which is rich in IPTG, bacteria can intake IPTG. IPTG is combined with LacI, then LacI cannot be bound to promoter Pgrac which is specially suppressed by LacI. Therefore, CⅠ repressor protein cannot generate as downstream cⅠ gene cannot transcribe regularly.
 +
As a result, P<sub>CⅠ</sub> recovers its activity and triggers the downstream transcription of GFP. By checking the expression of GFP, we could verify the feasibility of IPTG induction system, thus guaranteeing the successful culture of our engineered <i>Bacillus subtilis</i>.
  
 
Plasmid profile
 
Plasmid profile
https://2020.igem.org/wiki/images/e/ea/T--NAU-CHINA--composite-parts4.1.png
+
https://2020.igem.org/wiki/images/5/58/T--NAU-CHINA--composite-parts6.1.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 14:30, 16 October 2020

PliaG-B0034-lacI-Pgrac-B0034-CⅠ-PCⅠ-GFP-B0015

This composite part consists of promoter PliaG and Pgrac, lacI gene, CⅠ gene, GFP gene and some essential RBS and terminators. Promoter PliaG starts transcription process and its transcription production is LacI, which is regulated by IPTG. When bacteria is exposed to medium which is rich in IPTG, bacteria can intake IPTG. IPTG is combined with LacI, then LacI cannot be bound to promoter Pgrac which is specially suppressed by LacI. Therefore, CⅠ repressor protein cannot generate as downstream cⅠ gene cannot transcribe regularly. As a result, PCⅠ recovers its activity and triggers the downstream transcription of GFP. By checking the expression of GFP, we could verify the feasibility of IPTG induction system, thus guaranteeing the successful culture of our engineered Bacillus subtilis.

Plasmid profile T--NAU-CHINA--composite-parts6.1.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 1294
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2155