Difference between revisions of "Part:BBa K1111015"
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==Introduction==
 
==Introduction==
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This part is a Fusion protein made respectively of an Ice Nucleation Protein (INP), EYFP and a Streptavidin protein.
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This part is a Fusion protein made respectively of an Ice Nucleation Protein (INP), EYFP and a Streptavidin protein; which is an improved version of BBa_K1111014.
 +
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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[[File:EPF-Lausanne map.jpg||thumb|400px|center|Figure 1: reminder of BBa_K1111014 expected outcome]]
  
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<!-- Add more about the biology of this part here
 
 
===Usage and Biology===
 
===Usage and Biology===
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This Biobrick is designed to allow the INP to target streptavidin on the outer membrane of E.coli with EYFP as a reporter.
 +
Indeed EYFP fluorescence will assess of the construct expression and Immunostaining assay (as we did for BBa_K523013) will be used to show the localisation on the cell membrane.
 +
We could also target streptavidin with a specific antibody and perform biotin staining assays to see the functionnality of the streptavidin expressed.
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Our final goal is to attach biotynilated Gelatine nanoparticules to the E.coli cells thanks to the biotin-streptavidin interaction.
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K1111015 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1111015 SequenceAndFeatures</partinfo>

Revision as of 21:25, 28 October 2013

INP_EYFP_Streptavidin Fusion Protein

Introduction

This part is a Fusion protein made respectively of an Ice Nucleation Protein (INP), EYFP and a Streptavidin protein; which is an improved version of BBa_K1111014. 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: reminder of BBa_K1111014 expected outcome

Usage and Biology

This Biobrick is designed to allow the INP to target streptavidin on the outer membrane of E.coli with EYFP as a reporter. Indeed EYFP fluorescence will assess of the construct expression and Immunostaining assay (as we did for BBa_K523013) will be used to show the localisation on the cell membrane. We could also target streptavidin with a specific antibody and perform biotin staining assays to see the functionnality of the streptavidin expressed. Our final goal is to attach biotynilated Gelatine nanoparticules to the E.coli cells thanks to the biotin-streptavidin interaction.


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 2384
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1036
    Illegal AgeI site found at 2426
    Illegal AgeI site found at 2477
  • 1000
    COMPATIBLE WITH RFC[1000]