Difference between revisions of "Part:BBa K3328039"

 
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An integral part of IMPLY1 boolean calculation
 
An integral part of IMPLY1 boolean calculation
 
===Design===
 
===Design===
We combined the 3WJ switch and toehold switch to realize the IMPLY Boolean calculation. When no trigger expressed, this logic gate just likes a 3WJ switch. When trigger A expressed, the trigger will bind to the switch RNA. The binding allows for a branch migration process, exposing AUG and RBS for translation initiation. When trigger B expressed, the trigger will bind to 3WJ switch RNA. The resulting trigger–switch complex has a stable 3WJ structure that effectively sequesters the RBS and start codon within the loop and stem of the switch RNA, respectively, and strongly represses translation. When trigger A and B both expressed, the binding of trigger RNA to the toehold sequence allow the RNA polymerase binds to the former RBS and break open 3WJ stable hairpin.
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We combined the 3WJ switch and toehold switch to realize the IMPLY Boolean calculation. When no trigger is expressed, this logic gate just likes a 3WJ switch. When trigger A expressed, the trigger will bind to the switch RNA. The binding allows for a branch migration process, exposing AUG and RBS for translation initiation. When trigger B expressed, the trigger will bind to 3WJ switch RNA. The resulting trigger–switch complex has a stable 3WJ structure that effectively sequesters the RBS and the start codon within the loop and stem of the switch RNA, respectively, and strongly represses translation. When both trigger A and B are expressed, the binding of trigger RNA to the toehold sequence allows the RNA polymerase to bind to the former RBS and break open 3WJ stable hairpin.
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https://2020.igem.org/wiki/images/thumb/f/f7/T--OUC-China--design_lunbo_imply.jpg/799px-T--OUC-China--design_lunbo_imply.jpg
  
 
===Result===
 
===Result===
 
https://2020.igem.org/wiki/images/thumb/5/52/T--OUC-China--part_IMPLY1.jpg/800px-T--OUC-China--part_IMPLY1.jpg
 
https://2020.igem.org/wiki/images/thumb/5/52/T--OUC-China--part_IMPLY1.jpg/800px-T--OUC-China--part_IMPLY1.jpg
  
The left figure shows that when INPUT A=0, INPUT B=1, the fluorescence of GFP is low. And the fluorescence intensity of GFP was high in the other three groups. This corresponds to the situation described in the truth table on the right. INPUT A=1 means that aTc (0.25 mg/ml) is added, INPUT B=1 means that HSL (0.1 mg/ml) is added.
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The left figure shows that when INPUT A=0, INPUT B=1, the fluorescence of GFP is low. And the fluorescence intensity of GFP was high in the other three groups. This corresponds to the situation described in the truth table on the right. INPUT A=1 means that aTc (0.25 mg/mL) is added, INPUT B=1 means that HSL (0.1 mg/mL) is added. Error bar: SD (n=9).
 
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 12:02, 27 October 2020


triggerA of IMPLY gate (IMPLY1)

An integral part of IMPLY1 boolean calculation

Design

We combined the 3WJ switch and toehold switch to realize the IMPLY Boolean calculation. When no trigger is expressed, this logic gate just likes a 3WJ switch. When trigger A expressed, the trigger will bind to the switch RNA. The binding allows for a branch migration process, exposing AUG and RBS for translation initiation. When trigger B expressed, the trigger will bind to 3WJ switch RNA. The resulting trigger–switch complex has a stable 3WJ structure that effectively sequesters the RBS and the start codon within the loop and stem of the switch RNA, respectively, and strongly represses translation. When both trigger A and B are expressed, the binding of trigger RNA to the toehold sequence allows the RNA polymerase to bind to the former RBS and break open 3WJ stable hairpin.

799px-T--OUC-China--design_lunbo_imply.jpg

Result

800px-T--OUC-China--part_IMPLY1.jpg

The left figure shows that when INPUT A=0, INPUT B=1, the fluorescence of GFP is low. And the fluorescence intensity of GFP was high in the other three groups. This corresponds to the situation described in the truth table on the right. INPUT A=1 means that aTc (0.25 mg/mL) is added, INPUT B=1 means that HSL (0.1 mg/mL) is added. Error bar: SD (n=9).

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]