Difference between revisions of "Part:BBa K533003:Experience"
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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter | This experience page is provided so that any user may enter their experience using this part.<BR>Please enter | ||
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===Applications of BBa_K533003=== | ===Applications of BBa_K533003=== | ||
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+ | Like [[Part:BBa_I712667 | BBa_I712667]], it can specifically cleave proteins in the peptide. The difference is that it is expressed as a membrane protein, which enables it to cut target proteins in the media. | ||
+ | |||
+ | We tested function of the protease with OmpA-HIV protese site-SH3, our part [[Part:BBa_K533002 | BBa_K533002]]. | ||
+ | |||
+ | E. coli expressing OmpA-HIV protease site-SH3 were first mixed up in multi-proline mCherry, to give it the red fluorescence marker. After washing with distilled water twice, the remaining bacteria were mixed with E. coli expressing OmpA-HIV protease at room temperature for 30 minutes to release the cargo. | ||
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+ | After washing with distilled water twice, bacteria were fixed on a slip and stained with DAPI. | ||
+ | |||
+ | |||
+ | {| style="background: none; text-align:center;" align="center" | ||
+ | | Color Channel | ||
+ | | + E. coli expressing protease | ||
+ | | + E. coli without protease | ||
+ | |- | ||
+ | | mCherry | ||
+ | | [[Image:Thuexp_release_p.png|350px]] | ||
+ | | [[Image:Thuexp_release_n.png|350px]] | ||
+ | |- | ||
+ | | DAPI | ||
+ | | [[Image:Thuexp_release_p_d.png|350px]] | ||
+ | | [[Image:Thuexp_release_n_d.png|350px]] | ||
+ | |- | ||
+ | | merge | ||
+ | | [[Image:Thuexp_release_p_m.png|350px]] | ||
+ | | [[Image:Thuexp_release_n_m.png|350px]] | ||
+ | |} | ||
===User Reviews=== | ===User Reviews=== |
Revision as of 14:04, 5 October 2011
This experience page is provided so that any user may enter their experience using this part.
Please enter
how you used this part and how it worked out.
Applications of BBa_K533003
Like BBa_I712667, it can specifically cleave proteins in the peptide. The difference is that it is expressed as a membrane protein, which enables it to cut target proteins in the media.
We tested function of the protease with OmpA-HIV protese site-SH3, our part BBa_K533002.
E. coli expressing OmpA-HIV protease site-SH3 were first mixed up in multi-proline mCherry, to give it the red fluorescence marker. After washing with distilled water twice, the remaining bacteria were mixed with E. coli expressing OmpA-HIV protease at room temperature for 30 minutes to release the cargo.
After washing with distilled water twice, bacteria were fixed on a slip and stained with DAPI.
Color Channel | + E. coli expressing protease | + E. coli without protease |
mCherry | ||
DAPI | ||
merge |
User Reviews
UNIQ8358d49eab5c93a6-partinfo-00000000-QINU UNIQ8358d49eab5c93a6-partinfo-00000001-QINU