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Team:HUST-China/Project/LCD/Result
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<h2>Result</h2> | <h2>Result</h2> | ||
| - | <img src="https://static.igem.org/mediawiki/2012/4/45/Hust2012immunofluorescence.jpg"></img> | + | <img src="https://static.igem.org/mediawiki/2012/4/45/Hust2012immunofluorescence.jpg" width=700></img> |
Fig. 1 Immunofluorescence labeling of transformed cells: green fluorescence corresponds to cells labeled with Mouse Anti HA-Tag Polyclonal antibody and (Fluorescein (FITC)–conjugated Affinipure Goat Anti-Mouse IgG(H+L), and red fluorescence corresponds to cells labeled with Rabbit Anti Flag-Tag Polyclonal antibody and Goat Anti Rabbit RPE Polyclonal antibody. <br/> | Fig. 1 Immunofluorescence labeling of transformed cells: green fluorescence corresponds to cells labeled with Mouse Anti HA-Tag Polyclonal antibody and (Fluorescein (FITC)–conjugated Affinipure Goat Anti-Mouse IgG(H+L), and red fluorescence corresponds to cells labeled with Rabbit Anti Flag-Tag Polyclonal antibody and Goat Anti Rabbit RPE Polyclonal antibody. <br/> | ||
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We fused HA-tag with N-terminal of bglX gene on ZXFα-bglX vector. Label cells with Mouse Anti HA-Tag Polyclonal antibody and Fluorescein (FITC)–conjugated Affinipure Goat Anti-Mouse IgG(H+L). Result of flow cytometry analysis is as follows: fluorescence intensity x-mean value of Pichia pastoris with pPICZα empty vector is 0.306, with bglX-ZXFα is 23.4.<br/> | We fused HA-tag with N-terminal of bglX gene on ZXFα-bglX vector. Label cells with Mouse Anti HA-Tag Polyclonal antibody and Fluorescein (FITC)–conjugated Affinipure Goat Anti-Mouse IgG(H+L). Result of flow cytometry analysis is as follows: fluorescence intensity x-mean value of Pichia pastoris with pPICZα empty vector is 0.306, with bglX-ZXFα is 23.4.<br/> | ||
| - | <img src="https://static.igem.org/mediawiki/2012/3/3e/Hust2012Fc1.jpg"></img> | + | <img src="https://static.igem.org/mediawiki/2012/3/3e/Hust2012Fc1.jpg" width=300></img> |
| - | <img src="https://static.igem.org/mediawiki/2012/4/4e/Hust2012Fc2.jpg"></img><br/> | + | <img src="https://static.igem.org/mediawiki/2012/4/4e/Hust2012Fc2.jpg" width=300></img><br/> |
Fig. 2-1 Left: pPICZα negative control (x-mean: 0.306/1.17); Right: bglX (x-mean: 23.4/23.6)<br/> | Fig. 2-1 Left: pPICZα negative control (x-mean: 0.306/1.17); Right: bglX (x-mean: 23.4/23.6)<br/> | ||
<h3>Immunofluorescence analysis of endoxylanase surface display Pichia pastoris using flow cytometry.</h3> | <h3>Immunofluorescence analysis of endoxylanase surface display Pichia pastoris using flow cytometry.</h3> | ||
We fused HA-tag with N-terminal of xyn gene on ZXFα-xyn vector. Label cells with Mouse Anti HA-Tag Polyclonal antibody and Fluorescein (FITC)–conjugated Affinipure Goat Anti-Mouse IgG(H+L). Result of flow cytometry analysis is as follows: fluorescence intensity x-mean value of Pichia pastoris with pPICZα empty vector is 0.62, with xyn-ZXFα is 67.<br/> | We fused HA-tag with N-terminal of xyn gene on ZXFα-xyn vector. Label cells with Mouse Anti HA-Tag Polyclonal antibody and Fluorescein (FITC)–conjugated Affinipure Goat Anti-Mouse IgG(H+L). Result of flow cytometry analysis is as follows: fluorescence intensity x-mean value of Pichia pastoris with pPICZα empty vector is 0.62, with xyn-ZXFα is 67.<br/> | ||
| - | <img src="https://static.igem.org/mediawiki/2012/2/25/Hust2012Fc3.jpg"></img> | + | <img src="https://static.igem.org/mediawiki/2012/2/25/Hust2012Fc3.jpg" width=300></img> |
| - | <img src="https://static.igem.org/mediawiki/2012/b/bb/Hust2012Fc4.jpg"></img><br/> | + | <img src="https://static.igem.org/mediawiki/2012/b/bb/Hust2012Fc4.jpg" width=300></img><br/> |
Fig. 2-2 Left: pPICZα negative control (x-mean: 0.62/4.31); Right: xyn (x-mean: 67/67.6) | Fig. 2-2 Left: pPICZα negative control (x-mean: 0.62/4.31); Right: xyn (x-mean: 67/67.6) | ||
<h3>Immunofluorescence analysis of β-xylosidase Ruxyn surface display Pichia pastoris using flow cytometry.</h3> | <h3>Immunofluorescence analysis of β-xylosidase Ruxyn surface display Pichia pastoris using flow cytometry.</h3> | ||
We fused FLAG-tag with N-terminal of Ruxyn gene on Ruxyn-pPIC9k vector. Label cells with Rabbit Anti Flag-Tag Polyclonal antibody and Goat Anti Rabbit RPE Polyclonal antibody. Result of flow cytometry analysis is as follows: fluorescence intensity x-mean value of Pichia pastoris with pPIC9k empty vector is 1.14, with Ruxyn-pPIC9k is 8.56.<br/> | We fused FLAG-tag with N-terminal of Ruxyn gene on Ruxyn-pPIC9k vector. Label cells with Rabbit Anti Flag-Tag Polyclonal antibody and Goat Anti Rabbit RPE Polyclonal antibody. Result of flow cytometry analysis is as follows: fluorescence intensity x-mean value of Pichia pastoris with pPIC9k empty vector is 1.14, with Ruxyn-pPIC9k is 8.56.<br/> | ||
| - | <img src="https://static.igem.org/mediawiki/2012/6/6e/Fc5.jpg"></img> | + | <img src="https://static.igem.org/mediawiki/2012/6/6e/Fc5.jpg" width=300></img> |
| - | <img src="https://static.igem.org/mediawiki/2012/4/46/Fc6.jpg"></img><br/> | + | <img src="https://static.igem.org/mediawiki/2012/4/46/Fc6.jpg" width=300></img><br/> |
Fig. 2-3 Left: pPIC9k negative control (x-mean: 1.14/13.4); Right: Ruxyn (x-mean: 8.56/19.9) | Fig. 2-3 Left: pPIC9k negative control (x-mean: 1.14/13.4); Right: Ruxyn (x-mean: 8.56/19.9) | ||
<h3>Immunofluorescence analysis of endoglucanase cel5A surface display Pichia pastoris using flow cytometry.</h3> | <h3>Immunofluorescence analysis of endoglucanase cel5A surface display Pichia pastoris using flow cytometry.</h3> | ||
We fused FLAG-tag with N-terminal of cel5A gene on cel5A-pPIC9k vector. Label cells with Rabbit Anti Flag-Tag Polyclonal antibody and Goat Anti Rabbit RPE Polyclonal antibody. Result of flow cytometry analysis is as follows: fluorescence intensity x-mean value of Pichia pastoris with pPIC9k empty vector is 1.14, with cel5A-pPIC9k is 5.55.<br/> | We fused FLAG-tag with N-terminal of cel5A gene on cel5A-pPIC9k vector. Label cells with Rabbit Anti Flag-Tag Polyclonal antibody and Goat Anti Rabbit RPE Polyclonal antibody. Result of flow cytometry analysis is as follows: fluorescence intensity x-mean value of Pichia pastoris with pPIC9k empty vector is 1.14, with cel5A-pPIC9k is 5.55.<br/> | ||
| - | <img src="https://static.igem.org/mediawiki/2012/6/6e/Fc5.jpg"></img> | + | <img src="https://static.igem.org/mediawiki/2012/6/6e/Fc5.jpg" width=300></img> |
| - | <img src="https://static.igem.org/mediawiki/2012/b/bb/Fc8.jpg"></img> | + | <img src="https://static.igem.org/mediawiki/2012/b/bb/Fc8.jpg" width=300></img> |
<br/>Fig. 2-4 Left: pPIC9k negative control (x-mean: 1.14/13.4); Right: cel5A (x-mean: 5.55/16.5) | <br/>Fig. 2-4 Left: pPIC9k negative control (x-mean: 1.14/13.4); Right: cel5A (x-mean: 5.55/16.5) | ||
Revision as of 03:02, 27 September 2012
HUST CHINA
Result
Fig. 1 Immunofluorescence labeling of transformed cells: green fluorescence corresponds to cells labeled with Mouse Anti HA-Tag Polyclonal antibody and (Fluorescein (FITC)–conjugated Affinipure Goat Anti-Mouse IgG(H+L), and red fluorescence corresponds to cells labeled with Rabbit Anti Flag-Tag Polyclonal antibody and Goat Anti Rabbit RPE Polyclonal antibody. Immunofluorescence analysis of β-glucosidase surface display Pichia pastoris using flow cytometry.
We fused HA-tag with N-terminal of bglX gene on ZXFα-bglX vector. Label cells with Mouse Anti HA-Tag Polyclonal antibody and Fluorescein (FITC)–conjugated Affinipure Goat Anti-Mouse IgG(H+L). Result of flow cytometry analysis is as follows: fluorescence intensity x-mean value of Pichia pastoris with pPICZα empty vector is 0.306, with bglX-ZXFα is 23.4.
Fig. 2-1 Left: pPICZα negative control (x-mean: 0.306/1.17); Right: bglX (x-mean: 23.4/23.6)
Immunofluorescence analysis of endoxylanase surface display Pichia pastoris using flow cytometry.
We fused HA-tag with N-terminal of xyn gene on ZXFα-xyn vector. Label cells with Mouse Anti HA-Tag Polyclonal antibody and Fluorescein (FITC)–conjugated Affinipure Goat Anti-Mouse IgG(H+L). Result of flow cytometry analysis is as follows: fluorescence intensity x-mean value of Pichia pastoris with pPICZα empty vector is 0.62, with xyn-ZXFα is 67.
Fig. 2-2 Left: pPICZα negative control (x-mean: 0.62/4.31); Right: xyn (x-mean: 67/67.6)
Immunofluorescence analysis of β-xylosidase Ruxyn surface display Pichia pastoris using flow cytometry.
We fused FLAG-tag with N-terminal of Ruxyn gene on Ruxyn-pPIC9k vector. Label cells with Rabbit Anti Flag-Tag Polyclonal antibody and Goat Anti Rabbit RPE Polyclonal antibody. Result of flow cytometry analysis is as follows: fluorescence intensity x-mean value of Pichia pastoris with pPIC9k empty vector is 1.14, with Ruxyn-pPIC9k is 8.56.
Fig. 2-3 Left: pPIC9k negative control (x-mean: 1.14/13.4); Right: Ruxyn (x-mean: 8.56/19.9)
Immunofluorescence analysis of endoglucanase cel5A surface display Pichia pastoris using flow cytometry.
We fused FLAG-tag with N-terminal of cel5A gene on cel5A-pPIC9k vector. Label cells with Rabbit Anti Flag-Tag Polyclonal antibody and Goat Anti Rabbit RPE Polyclonal antibody. Result of flow cytometry analysis is as follows: fluorescence intensity x-mean value of Pichia pastoris with pPIC9k empty vector is 1.14, with cel5A-pPIC9k is 5.55.
Fig. 2-4 Left: pPIC9k negative control (x-mean: 1.14/13.4); Right: cel5A (x-mean: 5.55/16.5)
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