Device

Part:BBa_K533002

Designed by: ZHANG Yunxiao   Group: iGEM11_Tsinghua   (2011-09-29)
Revision as of 15:19, 5 October 2011 by Wishyx (Talk | contribs)

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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]


[edit]
Categories
//cds/membrane
//plasmid/expression/t7
Parameters
negative_regulatorstemperature, expressed below 20 centigrade
positive_regulatorsIPTG, 0.5mM