Difference between revisions of "Part:BBa K5121019"

 
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== Biology ==
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===Usage and Biology===
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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 — [Part:BBa_K2912000|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).
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K5121019 SequenceAndFeatures</partinfo>
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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:Part:BBa_K5121019:Design|design] page for more information.
  
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<span class='h3bb'><u>Sequence and Features</u></span>
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===Functional Parameters===
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<partinfo>BBa_K5121027 SequenceAndFeatures</partinfo>
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<partinfo>BBa_K5121019 parameters</partinfo>
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== References ==
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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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Revision as of 11:54, 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 — [Part:BBa_K2912000|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:Part:BBa_K5121019:Design|design] page for more information.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 803
    Illegal NheI site found at 865
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 745
  • 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.