Difference between revisions of "Part:BBa K1111012"
(→‎Usage and Biology)
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==Usage and Biology==
 
==Usage and Biology==
  
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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. Nanocapsules have a size comprised in a range from , which is quite big compared to a typical E:Coli cell size, we can therefore imagine attaching large proteins to these bacteria.
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This part was designed to express streptavidin on the outer membrane of E.coli in order to attach biotinylated nanoparticles to the cells. 
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Thus, it can be used in any experiment involving biotin-streptavidin to put cargos on bacteria.
  
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Revision as of 17:59, 2 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, their Kd is 10-15 M).


Usage and Biology

This part was designed to express streptavidin on the outer membrane of E.coli in order to attach biotinylated nanoparticles to the cells. Thus, it can be used in any experiment involving biotin-streptavidin to put cargos on bacteria.

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]