Difference between revisions of "Part:BBa K627009"

 
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<partinfo>BBa_K627009 short</partinfo>
 
<partinfo>BBa_K627009 short</partinfo>
  
 
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This BioBrick is a 2 parts fusion of pBAD arabinose-inducible induction system and the TEV protease. TEV protease is the common name for the 27 kDa catalytic domain of the Nuclear Inclusion a endopeptidase (NIa) encoded by the tobacco etch virus (TEV). TEV protease is a useful reagent for cleaving fusion proteins. It recognizes a linear epitope of the general form E-Xaa-Xaa-Y -Xaa-Q-(G/S), with cleavage occurring between Q and G or Q and S. In TEV protease the serine nucleophile of the conventional Ser-Asp-His triad is a cysteine instead. This probably explains why TEV protease is resistant to many commonly used protease inhibitors.
   
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At 37°C, the TEV protease forms inculsion body, which leads to an inactive form. Incubated at 30 °C, the protease is expressed as soluble type and is highly active. The induction system was amplified via PCR und fused via NgoMIV with the protease.
  
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
<partinfo>BBa_K627009 SequenceAndFeatures</partinfo>
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<partinfo>BBa_K627008 SequenceAndFeatures</partinfo>
  
  
 
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===Functional Parameters===
 
===Functional Parameters===
<partinfo>BBa_K627009 parameters</partinfo>
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<partinfo>BBa_K627008 parameters</partinfo>
 
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Revision as of 15:57, 21 September 2011

Arabinose inducible TEV protease

This BioBrick is a 2 parts fusion of pBAD arabinose-inducible induction system and the TEV protease. TEV protease is the common name for the 27 kDa catalytic domain of the Nuclear Inclusion a endopeptidase (NIa) encoded by the tobacco etch virus (TEV). TEV protease is a useful reagent for cleaving fusion proteins. It recognizes a linear epitope of the general form E-Xaa-Xaa-Y -Xaa-Q-(G/S), with cleavage occurring between Q and G or Q and S. In TEV protease the serine nucleophile of the conventional Ser-Asp-His triad is a cysteine instead. This probably explains why TEV protease is resistant to many commonly used protease inhibitors. At 37°C, the TEV protease forms inculsion body, which leads to an inactive form. Incubated at 30 °C, the protease is expressed as soluble type and is highly active. The induction system was amplified via PCR und fused via NgoMIV with the protease.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1239
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961
    Illegal SapI.rc site found at 1563