Difference between revisions of "Part:BBa K533002"

 
 
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__NOTOC__
 
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<partinfo>BBa_K533002 short</partinfo>
 
<partinfo>BBa_K533002 short</partinfo>
  
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Added HIV protease site between OmpA and SH3 for easier release.
 
  
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<!-- Add more about the biology of this part here
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===Usage and Biology===
 
===Usage and Biology===
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T7 promoter driven OmpA-HIV protease site-SH3 fusion protein expression cassette.
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This part is used for binding to target protein with multi-proline sequence. The target protein can then be efficiently released by HIV protease.
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{| style="background: none; text-align:center;" align="center"
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! Color Channel
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! E. coli with IPTG induction
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! E. coli without induction
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|-
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| mCherry
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| [[Image:Thuexp_bind_p.png|200px]]
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| [[Image:Thuexp_bind_n.png|200px]]
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|-
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| DAPI
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| [[Image:Thuexp_bind_p_d.png|200px]]
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| [[Image:Thuexp_bind_n_d.png|200px]]
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|-
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| merge
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| [[Image:Thuexp_bind_p_m.png|200px]]
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| [[Image:Thuexp_bind_n_m.png|200px]]
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|}
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The remaining 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.
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 +
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{| style="background: none; text-align:center;" align="center"
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! Color Channel
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! + E. coli expressing protease
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! + E. coli without protease
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|-
 +
| mCherry
 +
| [[Image:Thuexp_release_p.png|200px]]
 +
| [[Image:Thuexp_release_n.png|200px]]
 +
|-
 +
| DAPI
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| [[Image:Thuexp_release_p_d.png|200px]]
 +
| [[Image:Thuexp_release_n_d.png|200px]]
 +
|-
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| merge
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| [[Image:Thuexp_release_p_m.png|200px]]
 +
| [[Image:Thuexp_release_n_m.png|200px]]
 +
|}
  
 
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Latest revision as of 15:19, 5 October 2011

OmpA-HIV protease site-SH3



Usage and Biology

T7 promoter driven OmpA-HIV protease site-SH3 fusion protein expression cassette.

This part is used for binding to target protein with multi-proline sequence. The target protein can then be efficiently released by HIV protease.

Color Channel E. coli with IPTG induction E. coli without induction
mCherry Thuexp bind p.png Thuexp bind n.png
DAPI Thuexp bind p d.png Thuexp bind n d.png
merge Thuexp bind p m.png Thuexp bind n m.png

The remaining 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 Thuexp release p.png Thuexp release n.png
DAPI Thuexp release p d.png Thuexp release n d.png
merge Thuexp release p m.png Thuexp release n m.png

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 857
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]