Difference between revisions of "Part:BBa K1689012"
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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. <br/>
 
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. <br/>
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In our project, β-Lactamase is used to catalyze the hydrolysis of penicillin to penicillinoic acid, leading to current changes to be detected using A3 electrode system. We can dissect β-Lactamase between Gly196 and Leu198, for this is opposite to the active site; meanwhile we can acquire a more active and metabolically stable pattern by introducing a mutation M182T, which disrupts an inactive molten-globule intermediate of β-Lactamase.<br/>
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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.<br/>
 
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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Revision as of 15:04, 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


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]