Difference between revisions of "Part:BBa K1111014"
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<partinfo>BBa_K1111014 short</partinfo>
 
<partinfo>BBa_K1111014 short</partinfo>
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<!--INP sequence fused with Streptavidin (BBa_K283010) by gibson assembly-->
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INP sequence fused with Streptavidin (BBa_K283010) by gibson assembly
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==Introduction==
 
==Introduction==
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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.
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It is a fusion protein between an Ice Nucleation Protein (INP) and Streptavidin Alive made by Gibson Assembly.
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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).
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INP is an outer membrane protein foud in the genome of Pseudomonas syringae, that is recognize by the E.coli protein secretion machinery.  
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<!--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).-->
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[[File:EPF-Lausanne map.jpg||thumb|400px|center|Figure 1: outline of the expected outcome of the plasmid]]
  
 
==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.
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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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<br>Thus, it can be used in any experiment involving biotin-streptavidin to put cargos on bacteria.
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==Datasheet==
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For further Characterization, refer to the datasheet below.
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[[File:Team-EPF-Lausanne_Sheet1.jpg|thumb|270px|left|CharacterizationSheet Page1 ]]
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[[File: Team-EPF-Lausanne_Sheet2.jpg|thumb|270px|left|CharacterizationSheet Page2 ]]
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<br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br>
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<br><br>[[File: Team-EPF-Lausanne_Sheet.pdf|thumb|270px|left|PDF link ]]
  
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Thanks to the
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==<span class='h3bb'>Sequence and Features</span>==
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K1111014 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1111014 SequenceAndFeatures</partinfo>
  

Latest revision as of 18:19, 4 October 2013

Ice Nucleation Protein fused to Streptavidin BBa_K283010

Introduction

It is a fusion protein between an Ice Nucleation Protein (INP) and Streptavidin Alive made by Gibson Assembly. INP is an outer membrane protein foud in the genome of Pseudomonas syringae, that is recognize by the E.coli protein secretion machinery.

Figure 1: outline of the expected outcome of the plasmid

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.

Datasheet

For further Characterization, refer to the datasheet below.

CharacterizationSheet Page1
CharacterizationSheet Page2























File:Team-EPF-Lausanne Sheet.pdf

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 1649
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1036
    Illegal AgeI site found at 1691
    Illegal AgeI site found at 1742
  • 1000
    COMPATIBLE WITH RFC[1000]