Part:BBa_K1975008
Beta-Lactamase
This part contain a beta-lactamase. The function of this enzyme is to degrade beta-lactam and other beta-lactam antibiotics. The way the enzyme does this is by enzymatic cleavage of the beta-lactam ring. This cleavage will open the ring and the molecule will loose its antibacterial effect. This BioBrick consists only of the coding sequence, and does not contain any promotor. However, we can recommend to use the ampR promotor upstream this BioBrick.
Usage and Biology
Because of this activity, this enzyme is a very good reporter gene for Escherichia coli. When the E. coli cells contain this BioBrick the cells will survive to grow in media that contain beta-lactam. Cells without the BioBrick will die because beta-lactam will inhibit the cell wall biosynthesis making division of the cells impossible. This become especially useful after a transformation and positive cells will be screened. By using this BioBrick as reporter, only the cells that were transformed will survive on ampicillin containing plates.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Functional Parameters
Our results with this BioBricks showed that it was functional in Escherichia coli at both 50µg/mL and at 100µg/mL. This was validated by using E. coli that was transformed with a vector containing the gene and one that was transformed with only water. This clearly showed that the cells that were transformed with only water could not survive. This means that this BioBrick encodes ampicillin resistance, because it is the only gene that were available on the vector we used.
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