Difference between revisions of "Part:BBa K1689012"
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<partinfo>BBa_K1689012 short</partinfo> | <partinfo>BBa_K1689012 short</partinfo> | ||
− | N terminal of β-lactamase fused with dCas9<br/> | + | N-terminal fragment of β-lactamase fused with dCas9<br/> |
β-Lactamase (AmpR) 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 (AmpR) 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/> |
Revision as of 16:37, 27 September 2015
dCas9-Nlact
N-terminal fragment of β-lactamase fused with dCas9
β-Lactamase (AmpR) 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 [1]. We dissected β-Lactamase between Gly196 and Leu198 [2], 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 [2], which disrupts an inactive molten-globule intermediate of β-Lactamase.
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. Galarneau, A., Primeau, M., Trudeau, L. E., & Michnick, S. W. (2002). β-Lactamase protein fragment complementation assays as in vivo and in vitro sensors of protein–protein interactions. Nature biotechnology, 20(6), 619-622.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1150
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 4
Illegal BamHI site found at 3429 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]