Difference between revisions of "Part:BBa K5317014"

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===Usage and Biology===
 
===Usage and Biology===
  
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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). The CcpA inactivation impairs biofilm formation and restricts the incorporation of extracellular DNA (eDNA) into the biofilm matrix, while codY inactivation leads to a structured but robust biofilm bound to eDNA and intercellular adhesin polysaccharide (PIA) in ''S. aureus''(Bulock ''et al.'', 2022).
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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 bacterial protein to evaluate the interaction with the PknB kinase.
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=Cloning=
 
=Cloning=

Revision as of 14:32, 26 September 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 bacterial protein to evaluate the interaction with the PknB kinase.

Cloning

Theoretical Part Design

Placing the mRuby2 (K5317001) reporter gene behind ccpA allows for visualisation of expression of the protein of interest. Gene of interest was codon optimised for human cell lines.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

References

Bulock, L. L., Ahn, J., Shinde, D., Pandey, S., Sarmiento, C., Thomas, V. C., Guda, C., Bayles, K. W., & Sadykov, M. R. (2022). Interplay of CodY and CcpA in Regulating Central Metabolism and Biofilm Formation in Staphylococcus aureus. Journal of Bacteriology, 204(7), e00617-21. https://doi.org/10.1128/jb.00617-21

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