Difference between revisions of "Part:BBa K2450451:Design"

 
 
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===Design Notes===
 
===Design Notes===
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tba
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The spytag has been placed between OmpA and sfGFP. This is to ensure the insertion of OmpA into the membrane as we were concerned spytag at the N terminal end may disrupt the insertion of OmpA into the membrane. However, we were not sure if the isopeptide bond would be formed with the spytag in the centre of the protein. We therefore created another part with the opposite arrangement [BBa_K2450401], with the spytag at the N terminus of OmpA so we were less confident in the insertion of OmpA into the membrane, but more confident in the isopeptide bond. 
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sfGFP rather than GFP has been used as this part is being targeted to the outer membrane, and GFP cannot fold in the periplasmic space.
  
  
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===References===
 
===References===
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Alves, N. J., Turner, K. B., Medintz, I. L., & Walper, S. A. (2016). Protecting enzymatic function through directed packaging into bacterial outer membrane vesicles. Scientific Reports, 6(1), 24866. http://doi.org/10.1038/srep24866

Latest revision as of 13:17, 21 October 2017


OmpA SpyTag sfGFP tag


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 4
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 116


Design Notes

The spytag has been placed between OmpA and sfGFP. This is to ensure the insertion of OmpA into the membrane as we were concerned spytag at the N terminal end may disrupt the insertion of OmpA into the membrane. However, we were not sure if the isopeptide bond would be formed with the spytag in the centre of the protein. We therefore created another part with the opposite arrangement [BBa_K2450401], with the spytag at the N terminus of OmpA so we were less confident in the insertion of OmpA into the membrane, but more confident in the isopeptide bond.

sfGFP rather than GFP has been used as this part is being targeted to the outer membrane, and GFP cannot fold in the periplasmic space.


Source

tba

References

Alves, N. J., Turner, K. B., Medintz, I. L., & Walper, S. A. (2016). Protecting enzymatic function through directed packaging into bacterial outer membrane vesicles. Scientific Reports, 6(1), 24866. http://doi.org/10.1038/srep24866