Part:BBa_K3386000
ermC is a gene responsible for erythromycin resistance.
Usage and Biology
ermC encodes for a methyltransferase, which introduces dimethylation of the adenine residue in 23S rRNA at position 2085, thus preventing binding of macrolide and lincosamide antibiotics [1].
This part can be used to introduce resistance to macrolide antibiotics, such as erythromycin, which is a common selection marker. This gene is also appropriate for experiments, which rely on non-altering the concentration of antibiotics inside the cell or keeping the same chemical structure.
Characterization
In order to assess whether the ermC gene is functional, we have grown the cells for 20 h in various concentrations of antibiotic (10, 50 and 100 μg/ml). The antibiotics we chose were erythromycin, clarithromycin and spectinomycin. Erythromycin is our primer target for the biosensor we were constructing, since it is well described in literature, clarithromycin was chosen due to similar chemical structure, as we wanted to confirm the gene provides resistance for other macrolides. Spectinomycin was a negative control, since it belongs to a different group of antibiotics, but shares some of the structure with macrolide antibiotics: it also contains a lactone ring [2]. A positive control comprised cells grown in LB. A similar experiment was performed using non-transformed TOP10 cells, to ensure the difference in growth rates and survival was not a result of metabolic burden due to transformation with high copy number plasmid. All values were normalized. The lowest concentration of all of the antibiotics (10 ⲙg/ml) seem to not have a major effect on the cell growth. When it comes to cells grown in 50 and 100 ⲙg/ml, both transformed and non-transformed cells grown in spectinomycin showed no resistance towards this antibiotic. Media containing macrolide antibiotics seemed to only inhibit the growth of non-transformed cells (Fig. 1).
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]
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