Difference between revisions of "Part:BBa K5317014"

(Theoretical Part Design)
(Theoretical Part Design)
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===Theoretical Part Design===
 
===Theoretical Part Design===
Placing the mRuby2 reporter gene behind ccpA allows for visualisation of expression of the protein of interest. Gene of interest was codon optimised for human cell lines.
+
Placing the mRuby2 (<span class="plainlinks">[https://parts.igem.org/Part:BBa_K5317001 K5317001]</span>) 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===
 
===Sequence and Features===

Revision as of 14:26, 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). 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).


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