Difference between revisions of "Part:BBa K1111012"

 
Line 1: Line 1:
−
 
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K1111012 short</partinfo>
 
<partinfo>BBa_K1111012 short</partinfo>
  
−
INP sequence fused with Streptavidin Alive by gibson assembly.
+
=== Introduction ===
 +
 
 +
What it is : INP sequence fused with Streptavidin Alive by gibson assembly.
 +
The INP-streptavidin construct expresses streptavidin at the outer membrane of E.Coli. INP is an outer membrane protein foud in the genome of Pseudomonas syringae. It is under the control of Lac promoter and thus is constitutively expressed in E.Coli.
 +
The expression of this plasmid was tested by transforming cells and looking for microscopy at either of a fluorescent antibody again streptavidin or a fluorescent biotin (biotin and streptavidin have a strong affinity).
 +
 
  
−
<!-- Add more about the biology of this part here
 
 
===Usage and Biology===
 
===Usage and Biology===
  
−
<!-- -->
+
For us, the idea was to attach nanoparticles covered with biotin, using the strong affinity between both, but any experience implying biotin can be considered.
 +
 
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K1111012 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1111012 SequenceAndFeatures</partinfo>

Revision as of 20:20, 1 October 2013

Ice Nucleation Protein fused to Streptavidin Alive

Introduction

What it is : INP sequence fused with Streptavidin Alive by gibson assembly. The INP-streptavidin construct expresses streptavidin at the outer membrane of E.Coli. INP is an outer membrane protein foud in the genome of Pseudomonas syringae. It is under the control of Lac promoter and thus is constitutively expressed in E.Coli. The expression of this plasmid was tested by transforming cells and looking for microscopy at either of a fluorescent antibody again streptavidin or a fluorescent biotin (biotin and streptavidin have a strong affinity).


Usage and Biology

For us, the idea was to attach nanoparticles covered with biotin, using the strong affinity between both, but any experience implying biotin can be considered.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 1727
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1649
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 1727
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 1727
    Illegal NgoMIV site found at 1036
    Illegal AgeI site found at 1691
    Illegal AgeI site found at 1742
  • 1000
    COMPATIBLE WITH RFC[1000]