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<partinfo>BBa_I712667 AddReview 5</partinfo>
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<I>Yunxiao</I>
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We fused this protease to the C-terminal of OmpA and tested its cutting efficiency with a membrane protein containing its cleavage site.
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We induced OmpA-protease fusion protein with IPTG concentration of 0.1mM, 0.5mM and 1mM. After 12hours induction at 18 centigrade, protease activity were observed in all the samples.
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{| style="background: none; text-align:center;" align="center"
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| Color Channel
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| 0.1mM IPTG induction
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| 0.5mM IPTG induction
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| 1.0mM IPTG induction
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|-
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| mCherry
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| [[Image:Thuexp_release_0.1.png|250px]]
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| [[Image:Thuexp_release_0.5.png|250px]]
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| [[Image:Thuexp_release_1.0.png|250px]]
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|-
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| DAPI
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| [[Image:Thuexp_release_0.1_d.png|250px]]
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| [[Image:Thuexp_release_0.5_d.png|250px]]
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| [[Image:Thuexp_release_1.0_d.png|250px]]
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|-
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| merge
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| [[Image:Thuexp_release_0.1_m.png|250px]]
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| [[Image:Thuexp_release_0.5_m.png|250px]]
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| [[Image:Thuexp_release_1.0_m.png|250px]]
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|}
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''Protease cleavage will extinguish the red fluorescence from mCherry.''
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As is shown, the cleavage activity is highest at 0.1mM induction, suggesting the too high expression level might hinder protein folding.
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DAPI in the first set of data seems a bit diffusing, suggesting possible damage to the host cell.
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Latest revision as of 17:35, 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_I712667

User Reviews

UNIQ1dbd93be6925d920-partinfo-00000000-QINU

•••••

Yunxiao

We fused this protease to the C-terminal of OmpA and tested its cutting efficiency with a membrane protein containing its cleavage site.

We induced OmpA-protease fusion protein with IPTG concentration of 0.1mM, 0.5mM and 1mM. After 12hours induction at 18 centigrade, protease activity were observed in all the samples.

Color Channel 0.1mM IPTG induction 0.5mM IPTG induction 1.0mM IPTG induction
mCherry Thuexp release 0.1.png Thuexp release 0.5.png Thuexp release 1.0.png
DAPI Thuexp release 0.1 d.png Thuexp release 0.5 d.png Thuexp release 1.0 d.png
merge Thuexp release 0.1 m.png Thuexp release 0.5 m.png Thuexp release 1.0 m.png

Protease cleavage will extinguish the red fluorescence from mCherry.

As is shown, the cleavage activity is highest at 0.1mM induction, suggesting the too high expression level might hinder protein folding.

DAPI in the first set of data seems a bit diffusing, suggesting possible damage to the host cell.

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UNIQ1dbd93be6925d920-partinfo-00000002-QINU