Difference between revisions of "Part:BBa K5121019"
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== Biology == | == 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 — | + | 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 [ | + | 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 [Part:BBa_K5121019:Design|design] page for more information. |
<span class='h3bb'><u>Sequence and Features</u></span> | <span class='h3bb'><u>Sequence and Features</u></span> |
Revision as of 11:56, 2 October 2024
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 [Part:BBa_K5121019:Design|design] page for more information.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 803
Illegal NheI site found at 865 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 745
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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.