Difference between revisions of "Part:BBa K2623030"
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<partinfo>BBa_K2623030 short</partinfo>
 
<partinfo>BBa_K2623030 short</partinfo>
 
===Summary===
 
===Summary===
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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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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.  
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===Usage and Biology===
 +
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. We also constructed another form of BBa_K2623030 (NG9-1)-<partinfo>BBa_K2623028</partinfo> (NG8-1).The only difference is that the latter part links BBa_K2623022 (NG2) instead of NG4 with NG5. The fluorescence intensity of mRFP, we can compare and evaluate efficiency of SpyTag/SpyCather bioconjugation inside OMVs. Then the better one would be uesd to construct our final gene circuit-<partinfo>BBa_K2623032</partinfo>.
  
 
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Revision as of 14:01, 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.

Usage and Biology

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. We also constructed another form of BBa_K2623030 (NG9-1)-BBa_K2623028 (NG8-1).The only difference is that the latter part links BBa_K2623022 (NG2) instead of NG4 with NG5. The fluorescence intensity of mRFP, we can compare and evaluate efficiency of SpyTag/SpyCather bioconjugation inside OMVs. Then the better one would be uesd to construct our final gene circuit-BBa_K2623032.

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]