Difference between revisions of "Part:BBa K1689012"
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− | Reference: | + | 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. | + | 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. | + | 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/> |
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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.
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
- 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]