Difference between revisions of "Part:BBa K533003:Design"

 
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===Design Notes===
 
===Design Notes===
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Since HIV protease is a dimer, dimerization is a great concern to its function. Function of this protease is tested in BL21.
 
  
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We need a protease to release our binding cassette in our E. coli transportation system. HIV protease has a high specificity and a relatively simple structure, which suits our application well.
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We want to build an E. coli strain, expressing protease on its surface so that when it comes across another protein with the protease site, the protease can make the cut. Therefore, we fused the protein to the C-terminal of part of OmpA protein.
  
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The whole coding sequence is under T7 promoter and lacI repressor. When expressed in BL21 (DE3) strain, the best condition for expression is 0.5mM IPTG induced at 18 centigrade for 12 hours.
  
 
===Source===
 
===Source===

Latest revision as of 13:55, 5 October 2011

OmpA-HIV aspartyl protease


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 936
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We need a protease to release our binding cassette in our E. coli transportation system. HIV protease has a high specificity and a relatively simple structure, which suits our application well.

We want to build an E. coli strain, expressing protease on its surface so that when it comes across another protein with the protease site, the protease can make the cut. Therefore, we fused the protein to the C-terminal of part of OmpA protein.

The whole coding sequence is under T7 promoter and lacI repressor. When expressed in BL21 (DE3) strain, the best condition for expression is 0.5mM IPTG induced at 18 centigrade for 12 hours.

Source

HIV protease sequence is from former part BBa_I712667. OmpA sequence is from BL21(DE3) e. Coli strain.

References