Difference between revisions of "Part:BBa K1111015"

Revision as of 21:27, 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.
Our final goal is to attach biotynilated Gelatine nanoparticules to the E.coli cells thanks to the biotin-streptavidin interaction.
Thus, it can be used in any experiment involving biotin-streptavidin to put cargos on bacteria.

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]

@@ Line 8: / Line 8: @@
 [[File:EPF-Lausanne map.jpg||thumb|400px|center|Figure 1: reminder of BBa_K1111014 expected outcome]]
-===Usage and Biology===
+====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.
+<br>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.
+<br>Our final goal is to attach biotynilated Gelatine nanoparticules to the E.coli cells thanks to the biotin-streptavidin interaction.
-Our final goal is to attach biotynilated Gelatine nanoparticules to the E.coli cells thanks to the biotin-streptavidin interaction.
+<br>Thus, it can be used in any experiment involving biotin-streptavidin to put cargos on bacteria.