Difference between revisions of "Part:BBa K1111012"
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==Introduction== | ==Introduction== | ||
It is a fusion protein between an Ice Nucleation Protein (INP) and Streptavidin Alive made by Gibson Assembly. | It is a fusion protein between an Ice Nucleation Protein (INP) and Streptavidin Alive made by Gibson Assembly. | ||
| − | INP is an outer membrane protein | + | INP is an outer membrane protein found in the genome of Pseudomonas syringae, that is recognized by the E.coli protein secretion machinery, and thus also exported in that organism. |
| − | + | ||
| − | + | ||
<!--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).--> | <!--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== | ||
This part was designed to express streptavidin on the outer membrane of E.coli in order to attach biotinylated nanoparticles to the cells. | This part was designed to express streptavidin on the outer membrane of E.coli in order to attach biotinylated nanoparticles to the cells. | ||
| − | <br>Thus, it can be used in any experiment involving biotin-streptavidin to | + | <br>Thus, it can be used in any experiment involving biotin-streptavidin affinity (attachment of cargo is one example, but we could also think of purification etc...). |
| + | |||
| + | ==Datasheet== | ||
| + | For further Characterization, refer to the datasheet below. | ||
| + | [[File:Team-EPF-Lausanne_Sheet1.jpg|thumb|270px|left|CharacterizationSheet Page1 ]] | ||
| + | [[File: Team-EPF-Lausanne_Sheet2.jpg|thumb|270px|left|CharacterizationSheet Page2 ]] | ||
| + | <br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br> | ||
| + | <br><br>[[File: Team-EPF-Lausanne_Sheet.pdf|thumb|270px|left|PDF link ]] | ||
| − | <span class='h3bb'>Sequence and Features</span> | + | ==<span class='h3bb'>Sequence and Features</span>== |
<partinfo>BBa_K1111012 SequenceAndFeatures</partinfo> | <partinfo>BBa_K1111012 SequenceAndFeatures</partinfo> | ||
Latest revision as of 18:19, 4 October 2013
Ice Nucleation Protein fused to Streptavidin Alive
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 found in the genome of Pseudomonas syringae, that is recognized by the E.coli protein secretion machinery, and thus also exported in that organism.
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 affinity (attachment of cargo is one example, but we could also think of purification etc...).
Datasheet
For further Characterization, refer to the datasheet below.
File:Team-EPF-Lausanne Sheet.pdf
Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 1727
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1649
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 1727
- 25INCOMPATIBLE 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 - 1000COMPATIBLE WITH RFC[1000]