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Team:UCSF/HWP1
From 2011.igem.org
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<h3red>Hwp1</h3red> <p> | <h3red>Hwp1</h3red> <p> | ||
| - | <regulartext>HWP1 is a Candida albicans cell surface protein gene. It is used in Candida albicans to mediate tight binding to oral epithelial cells. Hwp1 is covalently linked to cell wall glucan through a remnant of its GPI anchor. (Eukaryot Cell. 2006 October; 5(10): 1604–1610.) Hwp1 is known to cause biofilm formation in vivo in C. albicans. | + | <regulartext>HWP1 is a <i>Candida albicans</i> cell surface protein gene. It is used in <i>Candida albicans</i> to mediate tight binding to oral epithelial cells. Hwp1 is covalently linked to cell wall glucan through a remnant of its GPI anchor. (Eukaryot Cell. 2006 October; 5(10): 1604–1610.) Hwp1 is known to cause biofilm formation in vivo in <i>C. albicans.</i> |
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| - | We cloned HWP1 into the plasmid pCTCON2. We used homologous recombination and transformed it into EBY100 Yeast. We used pCTCON2 because it has Aga2, which helps express HWP1 on the surface of the yeast cell. pCTCON2 also has the pGal1 promoter so HWP1 is only expressed in the presence of Galactose. A TRP marker is used as a selectable marker to tell the difference between yeast cells with the pCTCON2 plasmid and yeast cells without the pCTCON2 plasmid. S. cerevisiae will express HWP1 in the presence of Galactose and will not when it isn't present.<p><regulartext> | + | We cloned HWP1 into the plasmid pCTCON2. We used homologous recombination and transformed it into EBY100 Yeast. We used pCTCON2 because it has Aga2, which helps express HWP1 on the surface of the yeast cell. pCTCON2 also has the pGal1 promoter so HWP1 is only expressed in the presence of Galactose. A TRP marker is used as a selectable marker to tell the difference between yeast cells with the pCTCON2 plasmid and yeast cells without the pCTCON2 plasmid. <i>S. cerevisiae</i> will express HWP1 in the presence of Galactose and will not when it isn't present.<p><regulartext> |
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| - | <img align="left" style="margin-bottom:0px; width: 300px; margin-top:0px; padding:0;" src="http:// | + | <img align="left" style="margin-bottom:0px; width: 300px; margin-top:0px; padding:0;" src="http://dl.dropbox.com/u/24404809/Hwp1ctrl.jpg"> |
| - | <regulartext1> <b><br><BR><BR><BR><BR><br><BR><BR><BR><BR><BR><bR><BR><BR><BR><BR>Figure 1:</b> Control, non-induced | + | <regulartext1> <b><br><BR><BR><BR><BR><br><BR><BR><BR><BR><BR><bR><BR><BR><BR><BR>Figure 1:</b> Control, non-induced Hwp1 expressing yeast. Dispersed, no major |
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clustering. </regulartext1> | clustering. </regulartext1> | ||
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| - | <img align="left" style="margin-bottom:0px; width: 300px; margin-top:0px; padding:0;" src="http:// | + | <img align="left" style="margin-bottom:0px; width: 300px; margin-top:0px; padding:0;" src="http://dl.dropbox.com/u/24404809/Hwp1Gal1.jpg"> |
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<regulartext1> <b><br><BR><BR><BR><BR><br><BR><BR><BR><BR><bR><BR><BR><BR><BR>Figure 2:</b> Experiment, induced mgfp- | <regulartext1> <b><br><BR><BR><BR><BR><br><BR><BR><BR><BR><bR><BR><BR><BR><BR>Figure 2:</b> Experiment, induced mgfp- | ||
Latest revision as of 02:16, 29 September 2011
Figure 1: Control, non-induced Hwp1 expressing yeast. Dispersed, no major clustering.
Figure 2: Experiment, induced mgfp- 5 yeast. 3-D cluster of yeast cells growing around and on top of other cells.
