Composite
aGFP-SpAB

Part:BBa_K4071000

Designed by: María Calvo Noriega   Group: iGEM21_Stockholm   (2021-08-03)


B domain of Staphylococcal protein A fused to amilGFP chromoprotein

B domain of Staphylococcal protein A fused to amilGFP chromoprotein.

The part contains a strong constitutive promoter, RBS, and the amilGFP-protein A fusion protein.


Usage and Biology

Protein A is a well-known protein from Staphylococcus aureus, abbreviated as SpA. It contains five highly homologous Ig-binding domains in tandem: domains E, D, A, B, and C. SpA acts as a virulence factor thanks to these immunoglobulin-binding domains, which bind to the constant fragments (Fc) of IgG antibodies facilitating immune evasion. [1]

The domain B of protein A (SpAB) is a short domain of 252 amino acids that folds into three alfa-helixes connected by disordered regions, which confer a high degree of mobility to the helixes. [2][3]

Protein a.jpg

source: PDB

Protein A is commonly used in molecular biology, i.e. for Tandem affinity purification, or in biotechnological applications, i.e. Affibodies. It also has a role in therapy, since SpA-based immunoadsorption has been proposed as a treatment for patients suffering from autoimmune diseases as well as to reverse severe antibody-mediated rejection in organ transplantations. [4]


AmilGFP is a yellow chromoprotein commonly used reporter. Its bright yellow colour can be observed by the naked eye, but it can also be quantified using spectrophotometry (absorption maximum at 502 nm and emission maximum at 512 nm). [5]

AmilGFP.JPG

source: [6]


Since protein A is a promising target for S. aureus detection systems and anti-S.aureus therapy, our amilGFP - protein A fusion protein will be useful for iGEM teams working with S. aureus-related projects.


References

[1][2] http://pfam.xfam.org/family/PF02216

[4] https://www.uniprot.org/uniprot/Q70AB8

[4] J. Ren, ... L.-Y. Jia. Comprehensive Biotechnology (Second Edition), 2011

[5][6] http://parts.igem.org/Part:BBa_K592010


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 860
    Illegal BamHI site found at 778
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//cds
//cds/receptor
//cds/reporter/chromoprotein
//function/reporter/color
Parameters
color
protein