Difference between revisions of "Part:BBa K5317014"
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===Usage and Biology=== | ===Usage and Biology=== | ||
| − | The regulatory functions of CcpA are modulated by phosphorylation by serine/threonine kinases, which can affect its DNA-binding activity and thus its ability to regulate target genes. This phosphorylation-dependent mechanism enables ''S. aureus'' to adapt to different environmental conditions, thereby increasing its survivability and virulence (Liao ''et al.'', 2022). We used this | + | The regulatory functions of CcpA are modulated by phosphorylation by serine/threonine kinases, which can affect its DNA-binding activity and thus its ability to regulate target genes. This phosphorylation-dependent mechanism enables ''S. aureus'' to adapt to different environmental conditions, thereby increasing its survivability and virulence (Liao ''et al.'', 2022). We used this mechanism of ccpA to evaluate the interaction with the PknB kinase when stimulated with ß-lactam antibiotics. |
=Cloning= | =Cloning= | ||
===Theoretical Part Design=== | ===Theoretical Part Design=== | ||
| − | This part was codon | + | This part was codon-optimized and synthesized for expression in human cell lines. |
| − | < | + | ===Sequence and Features=== |
| + | <partinfo>BBa_K5317014 SequenceAndFeatures</partinfo> | ||
| − | + | =Characterization= | |
| − | + | The correct ccpA functionality was analyzed by composing a gene cassette that is placed downstream of the constitutive CMV promoter and fused with the reporter gene mRuby2 to assess by indication with ß-lactam antibiotics the ccpA localization based on the fluorescent signal. Please visit the <span class="plainlinks">[https://parts.igem.org/Part:BBa_K5317019 K5317019]</span> registry entry to view the results. | |
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| + | =References= | ||
Liao, X., Li, H., Guo, Y., Yang, F., Chen, Y., He, X., Li, H., Xia, W., Mao, Z.-W., & Sun, H. (2022). Regulation of DNA-binding activity of the Staphylococcus aureus catabolite control protein A by copper (II)-mediated oxidation. ''Journal of Biological Chemistry'', 298(3), 101587. https://doi.org/10.1016/j.jbc.2022.101587 | Liao, X., Li, H., Guo, Y., Yang, F., Chen, Y., He, X., Li, H., Xia, W., Mao, Z.-W., & Sun, H. (2022). Regulation of DNA-binding activity of the Staphylococcus aureus catabolite control protein A by copper (II)-mediated oxidation. ''Journal of Biological Chemistry'', 298(3), 101587. https://doi.org/10.1016/j.jbc.2022.101587 | ||
Latest revision as of 22:07, 1 October 2024
CcpA
Usage and Biology
The regulatory functions of CcpA are modulated by phosphorylation by serine/threonine kinases, which can affect its DNA-binding activity and thus its ability to regulate target genes. This phosphorylation-dependent mechanism enables S. aureus to adapt to different environmental conditions, thereby increasing its survivability and virulence (Liao et al., 2022). We used this mechanism of ccpA to evaluate the interaction with the PknB kinase when stimulated with ß-lactam antibiotics.
Cloning
Theoretical Part Design
This part was codon-optimized and synthesized for expression in human cell lines.
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]
Characterization
The correct ccpA functionality was analyzed by composing a gene cassette that is placed downstream of the constitutive CMV promoter and fused with the reporter gene mRuby2 to assess by indication with ß-lactam antibiotics the ccpA localization based on the fluorescent signal. Please visit the K5317019 registry entry to view the results.
References
Liao, X., Li, H., Guo, Y., Yang, F., Chen, Y., He, X., Li, H., Xia, W., Mao, Z.-W., & Sun, H. (2022). Regulation of DNA-binding activity of the Staphylococcus aureus catabolite control protein A by copper (II)-mediated oxidation. Journal of Biological Chemistry, 298(3), 101587. https://doi.org/10.1016/j.jbc.2022.101587