Difference between revisions of "Part:BBa K1689012"

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https://static.igem.org/mediawiki/parts/4/4a/Peking-speculation-Amp.png   
 
https://static.igem.org/mediawiki/parts/4/4a/Peking-speculation-Amp.png   
 
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Reference:
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Reference:<br/>
1. do Prado, T. M., Foguel, M. V., Gonçalves, L. M., & Maria del Pilar, T. S. (2015). β-Lactamase-based biosensor for the electrochemical determination of benzylpenicillin in milk. Sensors and Actuators B: Chemical, 210, 254-258.
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1. do Prado, T. M., Foguel, M. V., Gonçalves, L. M., & Maria del Pilar, T. S. (2015). β-Lactamase-based biosensor for the electrochemical determination of benzylpenicillin in milk. Sensors and Actuators B: Chemical, 210, 254-258.<br/>
2. Shekhawat, S. S., & Ghosh, I. (2011). Split-protein systems: beyond binary protein–protein interactions. Current opinion in chemical biology, 15(6), 789-797.
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2. Shekhawat, S. S., & Ghosh, I. (2011). Split-protein systems: beyond binary protein–protein interactions. Current opinion in chemical biology, 15(6), 789-797.<br/><br/>
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 16:30, 27 September 2015

dCas9-Nlact

N terminal of β-lactamase fused with dCas9

β-Lactamase(Amp) is produced by some bacteria, providing resistance against penicilins and so on. The β-Lactamase hydrolyzes β-lactam's conservative ring region, deactivating the antibiotic, and it can be use for the treatment of bacterial infection.
In our project, β-Lactamase is used to catalyze the hydrolysis of penicillin to penicillinoic acid. This redox reaction leads to current changes, which can be detected using A3 electrode system. We dissected β-Lactamase between Gly196 and Leu198, for they are on the opposite side of the active site. Moreover, we can enhance the activity and metabolically stability of β-Lactamase by introducing a mutation M182T, which disrupts an inactive molten-globule intermediate of β-Lactamase.
Peking-speculation-Amp.png


Reference:
1. do Prado, T. M., Foguel, M. V., Gonçalves, L. M., & Maria del Pilar, T. S. (2015). β-Lactamase-based biosensor for the electrochemical determination of benzylpenicillin in milk. Sensors and Actuators B: Chemical, 210, 254-258.
2. Shekhawat, S. S., & Ghosh, I. (2011). Split-protein systems: beyond binary protein–protein interactions. Current opinion in chemical biology, 15(6), 789-797.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1150
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 4
    Illegal BamHI site found at 3429
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]