Difference between revisions of "Part:BBa K533001:Design"

 
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===Design Notes===
 
===Design Notes===
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High expression efficiency as well as inducible expression is most desirable when considering a coding sequence.
 
  
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High expression efficiency as well as inducible expression is most desirable when considering a coding sequence. We hence chose T7 promoter combined with lac repressor binding site to drive strong but inducible expression.
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OmpA is a common device for anchoring a protein onto the outer membrane of E. coli. Nonetheless, it's unnecessary to use the full length OmpA, only a short part in the N-terminal is sufficient. We chose the N-terminal 180 amino acids in reference to the previous part,  [Part:BBa_K103006  BBa_K103006]. We used more amino acids in OmpA than the previous part, with the intention that the extra amino acids can function as a linker.
  
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SH3 is a common protein domain in adapters, specifically binding to multi-proline sequence. We chose SH3 for our binding module, as it specifically binds to an unmodified peptide. Thus it offers an opportunity to add this peptide as a short tag onto any protein of desire, significantly widening the range of proteins that can be transported by our system.
  
 
===Source===
 
===Source===

Revision as of 13:21, 5 October 2011

OmpA-SH3


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 815
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

High expression efficiency as well as inducible expression is most desirable when considering a coding sequence. We hence chose T7 promoter combined with lac repressor binding site to drive strong but inducible expression.

OmpA is a common device for anchoring a protein onto the outer membrane of E. coli. Nonetheless, it's unnecessary to use the full length OmpA, only a short part in the N-terminal is sufficient. We chose the N-terminal 180 amino acids in reference to the previous part, [Part:BBa_K103006 BBa_K103006]. We used more amino acids in OmpA than the previous part, with the intention that the extra amino acids can function as a linker.

SH3 is a common protein domain in adapters, specifically binding to multi-proline sequence. We chose SH3 for our binding module, as it specifically binds to an unmodified peptide. Thus it offers an opportunity to add this peptide as a short tag onto any protein of desire, significantly widening the range of proteins that can be transported by our system.

Source

OmpA is derived from the genomic DNA of BL21 (DE3) E. Coli strain while SH3 is from human Grb2.

References