Coding
RebA

Part:BBa_K5121019

Designed by: Carlo Famularo   Group: iGEM24_Sydney-Australia   (2024-09-28)
Revision as of 12:03, 2 October 2024 by Mdepaulis800 (Talk | contribs)

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rebA C-teriminal LPETG

Biology

RebA encodes an 18 kDa protein which forms one of the main structural components of type 51 R bodies. This part is based on the wildtype rebA part — BBa_K2912000 — uploaded by SZU-China in 2019, however has been modified to be compatible with sortase A-mediated conjugation. Sortase A is a bacterial transpeptidase enzyme that ligates C-terminal LPXTG motifs with N-terminal polyglycine motifs (Theile et al., 2013).

Our group added an LPETG peptide sequence to the C-terminus of the rebA monomer. This modification facilitates conjugation of proteins/small molecules containing an N-terminal polyglycine motif to the C-terminus of rebA in the presence of sortase A. Using this approach, we conjugated the fluorescent protein mNeonGreen with an N-terminal polyglycine sequence (BBa_K5121012) to R-bodies containing the modified rebA monomer. See the design page for more information.

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

Theile, C. S., Witte, M. D., Blom, A. E. M., Kundrat, L., Ploegh, H. L., & Guimaraes, C. P. (2013). Site-specific N-terminal labeling of proteins using sortase-mediated reactions. Nature Protocols, 8(9), 1800–1807.

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