Difference between revisions of "Part:BBa K4088892"
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N-terminal of Npu DnaE intein.
 
N-terminal of Npu DnaE intein.
 
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You can find C-terminal of Npu DnaE intein there - <partinfo>BBa_K4088893</partinfo>
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You can find C-terminal of Npu DnaE intein there - <partinfo>BBa_K4088893</partinfo>.
 
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Intein is a segment of a protein that can self-catalytically excised out and ligate the remaining parts of the protein (N- and C-exteins) with a peptide bond <ref>Anraku, Y., Mizutani, R. and Satow, Y. (2005), Protein Splicing: Its Discovery and Structural Insight into Novel Chemical Mechanisms. IUBMB Life, 57: 563-574. https://doi.org/10.1080/15216540500215499</ref>.
 
Intein is a segment of a protein that can self-catalytically excised out and ligate the remaining parts of the protein (N- and C-exteins) with a peptide bond <ref>Anraku, Y., Mizutani, R. and Satow, Y. (2005), Protein Splicing: Its Discovery and Structural Insight into Novel Chemical Mechanisms. IUBMB Life, 57: 563-574. https://doi.org/10.1080/15216540500215499</ref>.
 
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For this part we used the Npu DnaE intein. This intein is identified as a naturally occurring split mini-intein in Synechocystis sp. PCC6803 <ref>Wu, H., Hu, Z., & Liu, X. Q. (1998). Protein trans-splicing by a split intein encoded in a split DnaE gene of Synechocystis sp. PCC6803. Proceedings of the National Academy of Sciences of the United States of America, 95(16), 9226–9231. https://doi.org/10.1073/pnas.95.16.9226</ref>. Its first amino acid is cysteine which is important for protein trans-splicing to take place.
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For this part we used the Npu DnaE intein. DnaE - alpha subunit of the DNA polymerase III intein. This intein is identified as a naturally occurring split intein in Nostoc punctiforme <ref>Oeemig JS, Aranko AS, Djupsjöbacka J, Heinämäki K, Iwaï H. Solution structure of DnaE intein from Nostoc punctiforme: structural basis for the design of a new split intein suitable for site-specific chemical modification. FEBS Lett. 2009 May 6;583(9):1451-6. https://doi.org/10.1016/j.febslet.2009.03.058</ref>. Its first amino acid is cysteine which is important for protein trans-splicing to take place.
 
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This part we used in the сomposite system - N-terminal fragment of β-lactamase fused with dCas13a and N-terminal intein (<partinfo>BBa_K4088891</partinfo>).
 
This part we used in the сomposite system - N-terminal fragment of β-lactamase fused with dCas13a and N-terminal intein (<partinfo>BBa_K4088891</partinfo>).

Latest revision as of 22:09, 21 October 2021


DnaE - N

N-terminal of Npu DnaE intein.

You can find C-terminal of Npu DnaE intein there - BBa_K4088893.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 260
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]





Usage and Biology

Intein
Intein is a segment of a protein that can self-catalytically excised out and ligate the remaining parts of the protein (N- and C-exteins) with a peptide bond [1].
For this part we used the Npu DnaE intein. DnaE - alpha subunit of the DNA polymerase III intein. This intein is identified as a naturally occurring split intein in Nostoc punctiforme [2]. Its first amino acid is cysteine which is important for protein trans-splicing to take place.

This part we used in the сomposite system - N-terminal fragment of β-lactamase fused with dCas13a and N-terminal intein (BBa_K4088891).



References

  1. ↑ Anraku, Y., Mizutani, R. and Satow, Y. (2005), Protein Splicing: Its Discovery and Structural Insight into Novel Chemical Mechanisms. IUBMB Life, 57: 563-574. https://doi.org/10.1080/15216540500215499
  2. ↑ Oeemig JS, Aranko AS, Djupsjöbacka J, Heinämäki K, Iwaï H. Solution structure of DnaE intein from Nostoc punctiforme: structural basis for the design of a new split intein suitable for site-specific chemical modification. FEBS Lett. 2009 May 6;583(9):1451-6. https://doi.org/10.1016/j.febslet.2009.03.058