Difference between revisions of "Part:BBa K4623003"
 
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<partinfo>BBa_K4623003 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K4623003 SequenceAndFeatures</partinfo>
  
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References:
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[1]Lucas M, González-Pérez B, Cabezas M, Moncalian G, Rivas G, de la Cruz F. Relaxase DNA binding and cleavage are two distinguishable steps in conjugative DNA processing that involve different sequence elements of the nic site. J Biol Chem. 2010 Mar 19;285(12):8918-26. doi: 10.1074/jbc.M109.057539. Epub 2010 Jan 8. PMID: 20061574; PMCID: PMC2838313.
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[2]Sagredo, Sandra et al. “Orthogonal Protein Assembly on DNA Nanostructures Using Relaxases.” *Angewandte Chemie (International ed. in English)* vol. 55,13 (2016): 4348-52. doi:10.1002/anie.201510313.
  
 
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Latest revision as of 12:37, 11 October 2023


TrwC, a protein linker to covalently link ssDNA 5' end

TrwC is a type of relaxase, which participates in bacterial DNA binding and horizontal transfer of genomic material between bacteria​ [1]​. In vitro, it can specifically recognize the nic sequence (GGTGCGTATTGTCT^ATAGC) on ssDNA, causing it to break, and covalently bind to the ssDNA.

Therefore, TrwC relaxase possesses sequence specificity, orthogonality, and the ability to form a covalent bond with standard oligonucleotides. This provides a potential method in our project design that may be superior to existing protein-bound DNA nanostructures. The linkage of TrwC with ssDNA in vitro depends on Mg2+, so the reaction can be initiated by adding Mg2+ in the mixed system. Conversely, adding EDTA can terminate the reaction and yield DNA-protein complexes [2]​.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 286
    Illegal PstI site found at 604
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 286
    Illegal PstI site found at 604
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 286
    Illegal PstI site found at 604
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 286
    Illegal PstI site found at 604
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 409
    Illegal SapI.rc site found at 124

References:

[1]Lucas M, González-Pérez B, Cabezas M, Moncalian G, Rivas G, de la Cruz F. Relaxase DNA binding and cleavage are two distinguishable steps in conjugative DNA processing that involve different sequence elements of the nic site. J Biol Chem. 2010 Mar 19;285(12):8918-26. doi: 10.1074/jbc.M109.057539. Epub 2010 Jan 8. PMID: 20061574; PMCID: PMC2838313.

[2]Sagredo, Sandra et al. “Orthogonal Protein Assembly on DNA Nanostructures Using Relaxases.” *Angewandte Chemie (International ed. in English)* vol. 55,13 (2016): 4348-52. doi:10.1002/anie.201510313.