Difference between revisions of "Part:BBa K1323011"
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| − | The construct (erythromycin cds in pUC57) was electroporated into s.epidermidis. Three colonies were streak purified and diagnostic restriction digests were done to confirm that the insert was present. Overnight TSB cultures were supplemented with 10 | + | The construct (erythromycin cds in pUC57) was electroporated into s.epidermidis. Three colonies were streak purified and diagnostic restriction digests were done to confirm that the insert was present. Overnight TSB cultures were supplemented with 10 µg/mL erythromycin, and the inoculation was done in triplicate. The negative control for this experiment was ''S.epidermidis'' (ATCC12228), it lacks erythromycin resistance but has chloramphenicol resistance. 10 µL of the overnight cultures were used to inoculate fresh TSB that was supplemented with 10, 50, or 100 µg/mL of erythromycin and chloramphenicol. The negative growth control was 75 µg/mL of kanamycin and the positive growth control was TSB without antibiotic. The samples were incubated at 37 ͒C, shaking at 200 rpm. The OD600 was read after 6 hours of incubation. |
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
<partinfo>BBa_K1323011 SequenceAndFeatures</partinfo> | <partinfo>BBa_K1323011 SequenceAndFeatures</partinfo> | ||
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| + | == References == | ||
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| + | Catchpole, I., Davies, T. A., and Dyke, K. G. H. (1988). The nucleotide sequence of Staphylococcus aureus plasmid pT48 conferring inducible macrolide-lincosamide-streptogramin B resistance and comparison with similar plasmids expressing constitutive resistance. Journal of General Microbiology 134, 697-709. | ||
Revision as of 04:51, 17 October 2014
Erythromycin resistance coding sequence
This coding sequence is for the antibiotic, erythromycin. This part is PCR amplified from ermM cassette synthesized by Biobasic. This ermM cassette is originally found in S.epidermidis plasmid pNE131 which confers resistance to macrolide-lincosamide-streptogramin B group antibiotics including erythromycin (Catchpole et al., 1988). Working concentration for this antibiotic is 10 ug/mL in S.epidermidis/S.aureus hosts.
The construct (erythromycin cds in pUC57) was electroporated into s.epidermidis. Three colonies were streak purified and diagnostic restriction digests were done to confirm that the insert was present. Overnight TSB cultures were supplemented with 10 µg/mL erythromycin, and the inoculation was done in triplicate. The negative control for this experiment was S.epidermidis (ATCC12228), it lacks erythromycin resistance but has chloramphenicol resistance. 10 µL of the overnight cultures were used to inoculate fresh TSB that was supplemented with 10, 50, or 100 µg/mL of erythromycin and chloramphenicol. The negative growth control was 75 µg/mL of kanamycin and the positive growth control was TSB without antibiotic. The samples were incubated at 37 ͒C, shaking at 200 rpm. The OD600 was read after 6 hours of incubation.
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]
References
Catchpole, I., Davies, T. A., and Dyke, K. G. H. (1988). The nucleotide sequence of Staphylococcus aureus plasmid pT48 conferring inducible macrolide-lincosamide-streptogramin B resistance and comparison with similar plasmids expressing constitutive resistance. Journal of General Microbiology 134, 697-709.