Difference between revisions of "Part:BBa K3805238:Design"
 
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===Design Notes===
 
===Design Notes===
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During syhthesis, we need to note that the volumn of replaced amino-acids should not be too large, for a large protein fragment will have a negative effect on functionating of receptor, causing the low binding ability of anti-mCherry pmrB and mChrrey. Hence, we replaced 131 of total 166 amino-acids, approximately half binding domain of anti-mCherry.
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During syhthesis, we need to note that the volumn of replaced amino-acids should not be too large, for a large protein fragment will have a negative effect on functionate of receptor, causing the low binding ability of anti-mCherry pmrB and mChrrey. Hence, we replaced 99 of total 166 amino-acids, approximately half binding domain of anti-mCherry.
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[[File:PmrB.png|center|thumb|400px|Figure 1: Anti-mCherry pmrB]]
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[[File:Anti-mCherry PmrB.png|center|thumb|400px|Figure 1: Anti-mCherry pmrB]]
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===Source===
 
===Source===
  
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Due to the specifity and limitation of pmrBAC system, the Iron-binding motif of pmrB was replaced with anti-mCherry sequence through synthesis.  
+
Due to the specifity and limitation of pmrBAC system, the Iron-binding motif of pmrB is replaced with anti-mCherry sequence through synthesis.  
  
 
===References===
 
===References===

Latest revision as of 23:24, 21 October 2021


anti-mCherry pmrB


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]


Design Notes

During syhthesis, we need to note that the volumn of replaced amino-acids should not be too large, for a large protein fragment will have a negative effect on functionate of receptor, causing the low binding ability of anti-mCherry pmrB and mChrrey. Hence, we replaced 99 of total 166 amino-acids, approximately half binding domain of anti-mCherry.

Figure 1: Anti-mCherry pmrB

Source

Due to the specifity and limitation of pmrBAC system, the Iron-binding motif of pmrB is replaced with anti-mCherry sequence through synthesis.

References

[1]Liang H, Deng X, Bosscher M, Ji Q, Jensen MP, He C. Engineering bacterial two-component system PmrA/PmrB to sense lanthanide ions. J Am Chem Soc. 2013 Feb 13;135(6):2037-9. doi: 10.1021/ja312032c. Epub 2013 Feb 1. PMID: 23350529. [2]Wösten MM, Groisman EA. Molecular characterization of the PmrA regulon. J Biol Chem. 1999 Sep 17;274(38):27185-90. doi: 10.1074/jbc.274.38.27185. PMID: 10480935. [3]Roland, K. L.; Martin, L. E.; Esther, C. R.; Spitznagel, J. K. J. Bacteriol. 1993, 175, 4154−4164. [4]Shafer, W. M.; Casey, S. G.; Spitznagel, J. K. Infect. Immun. 1984, 43, 834−838. [5]Wosten, M. M.; Kox, L. F.; Chamnongpol, S.; Soncini, F. C.; Groisman, E. A. Cell 2000, 103, 113−125.