Difference between revisions of "Part:BBa K627011"
Line 1: Line 1:
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K627011 short</partinfo><br>
 
<partinfo>BBa_K627011 short</partinfo><br>
−
This BioBrick is a 2 part fusion of arabinose-inducible induction system and the HRV 14_3C protease. The recombinant type 14_3C protease from human rhinovirus (HRV 3C) recognizes the same cleavage site as the native enzyme: LeuGluValLeuPheGln↓GlyPro.<br>
+
 
−
The small, 22 kDa size of the protease got its optimal activity at 4°C but is still very active at 37°C. It is commonly used for an easy tag removal after the purification of recombinant proteins carrying his-tag. The 14_3C works with a catalytic triade, containing the amino acid residues Ser-Asp-His at its active site.<br>
+
====Introduction====
−
The arabinose-inducible induction system was amplified via PCR from the pBAD_iGEMexpress vector and fused via NgoMIV with the HRV 14_3C protease. An effective induction can be accomplished with 2 mM up to 25 mM arabinose.<br>
+
This BioBrick is a 2 part fusion of arabinose-inducible induction system and the HRV 14_3C protease.
 +
====General Information of the single sub parts====
 +
=====HRV 14_3C=====
 +
The recombinant type 14_3C protease from human rhinovirus (HRV 3C) recognizes the same cleavage site as the native enzyme: LeuGluValLeuPheGln/GlyPro.<br>
 +
The 22 kDa protease got its optimal activity at 4°C but also shows a high cleavage rate at 37°C. The 14_3C works with a catalytic triade, containing the amino acid residues Ser-Asp-His at its active site.<br>
 +
=====Arabinose-inducible induction system=====
 +
The arabinose-inducible induction system was amplified via PCR from the pBAD_iGEMexpress vector. It gots a very low expression rate without induction by arabinose. For the induction concentration between 2 mM up to 50 mM arabinose can be used to get an high expression rate.
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 23:06, 21 September 2011

Fusion part of pBAD arabinose-inducible induction system and the HRV 14_3C protease

Introduction

This BioBrick is a 2 part fusion of arabinose-inducible induction system and the HRV 14_3C protease.

General Information of the single sub parts

HRV 14_3C

The recombinant type 14_3C protease from human rhinovirus (HRV 3C) recognizes the same cleavage site as the native enzyme: LeuGluValLeuPheGln/GlyPro.
The 22 kDa protease got its optimal activity at 4°C but also shows a high cleavage rate at 37°C. The 14_3C works with a catalytic triade, containing the amino acid residues Ser-Asp-His at its active site.

Arabinose-inducible induction system

The arabinose-inducible induction system was amplified via PCR from the pBAD_iGEMexpress vector. It gots a very low expression rate without induction by arabinose. For the induction concentration between 2 mM up to 50 mM arabinose can be used to get an high expression rate.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1523
    Illegal BglII site found at 1711
    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