Difference between revisions of "Part:BBa K2623030"

 
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<partinfo>BBa_K2623030 short</partinfo>
 
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===Summary===
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Under the induction of IPTG, this part can code OmpA-SpyTag and SpyCatcher-L7Ae-RFP to be encapsulated into OMVs through the isopeptide bond formed between SpyTag/SpyCatcher.
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This part is a composite part constructed by linking BBa_K2623024 and BBa_K2623025 together. BBa_K2623024 can express OmpA inserted with SpyTag (ST) at its C-termini.  BBa_K2623025 can express archaeal ribosomal protein protein L7Ae inserted with SpyCatcher (SC) at its N-termini and mRFP1 at its C-termini under the induction of IPTG at 0.5 mM concentration (Figure 1). Though we have revealed the OmpA-ST inside OMVs (BBa_K2623021), we are inclined to figure out the ST/SC bioconjugation inside OMVs. Once IPTG is added, L7Ae could be expressed with SpyCatcher and RFP modification and then be encapsulated into OMVs through ST/SC conjugation. OMVs labeled with RFP will be detected through HSFCM (Figure 2).  
  
 
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<span class='h3bb'>Sequence and Features</span>
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<span clas='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K2623030 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2623030 SequenceAndFeatures</partinfo>
  

Revision as of 13:20, 13 October 2018


A composite part of OmpA-SpyTag and SpyCatcher-L7Ae-mRFP1

Summary

This part is a composite part constructed by linking BBa_K2623024 and BBa_K2623025 together. BBa_K2623024 can express OmpA inserted with SpyTag (ST) at its C-termini. BBa_K2623025 can express archaeal ribosomal protein protein L7Ae inserted with SpyCatcher (SC) at its N-termini and mRFP1 at its C-termini under the induction of IPTG at 0.5 mM concentration (Figure 1). Though we have revealed the OmpA-ST inside OMVs (BBa_K2623021), we are inclined to figure out the ST/SC bioconjugation inside OMVs. Once IPTG is added, L7Ae could be expressed with SpyCatcher and RFP modification and then be encapsulated into OMVs through ST/SC conjugation. OMVs labeled with RFP will be detected through HSFCM (Figure 2).

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal NheI site found at 1965
    Illegal NheI site found at 1988
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 65
    Illegal AgeI site found at 3122
    Illegal AgeI site found at 3234
  • 1000
    COMPATIBLE WITH RFC[1000]