Composite

Part:BBa_K5317018

Designed by: Lisa Marie de Sousa Miranda   Group: iGEM24_Hannover   (2024-09-15)
Revision as of 14:50, 26 September 2024 by Vanessa09 (Talk | contribs) (→‎Usage and Biology)


CMV-EGFP-PknB

Usage and Biology

PknB is a eukaryote-like serine/threonine kinase in Staphylococcus aureus that plays an important role in the bacterial response to antibiotics, particularly beta-lactams, via its PASTA domain (Stehle et al.,2012). PknB is a membrane-localized protein consisting of an N-terminal cytosolic kinase domain, a central transmembrane segment and three C-terminal extracellular PASTA domains. The PASTA (penicillin-binding protein and serine/threonine kinase-associated) domain plays a critical role in the recognition and binding of beta-lactam antibiotics (Stehle et al.,2012). Upon binding these compounds, the PASTA domain initiates a signaling cascade by inducing autophosphorylation of the N-terminal kinase domain. This activation leads to the initiation of downstream signaling pathways (Cheung et al.,2010). In S. aureus, this mechanism is critical for early detection of antibiotics and helps the bacteria adapt to antibiotic stress (Sauer et al.,2018).

The composite part includes the upstream positioned reporter gene EGFP (K3338006) to charactarize the PknB regarding it's cellular localization pre and post antibiotics stimulation.

Cloning

Theoretical Part Design

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 2596
    Illegal SpeI site found at 3259
    Illegal PstI site found at 2021
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 3259
    Illegal PstI site found at 2021
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2271
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 2596
    Illegal SpeI site found at 3259
    Illegal PstI site found at 2021
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 2596
    Illegal SpeI site found at 3259
    Illegal PstI site found at 2021
  • 1000
    COMPATIBLE WITH RFC[1000]


References

Pensinger, D. A., Schaenzer, A. J., & Sauer, J. D. (2018). Do Shoot the Messenger: PASTA Kinases as Virulence Determinants and Antibiotic Targets. Trends in microbiology, 26(1), 56–69. https://doi.org/10.1016/j.tim.2017.06.010

Rakette S, Donat S, Ohlsen K, Stehle T (2012) Structural Analysis of Staphylococcus aureus Serine/Threonine Kinase PknB. PLOS ONE 7(6): e39136. https://doi.org/10.1371/journal.pone.0039136

Tamber, S., Schwartzman, J., & Cheung, A. L. (2010). Role of PknB kinase in antibiotic resistance and virulence in community-acquired methicillin-resistant Staphylococcus aureus strain USA300. Infection and immunity, 78(8), 3637–3646. https://doi.org/10.1128/IAI.00296-10

[edit]
Categories
Parameters
None