Difference between revisions of "Part:BBa K644001"
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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. | 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. | ||
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Revision as of 07:50, 22 October 2020
Candida albicans hyphal wall protein 1 (HWP1) gene
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.
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.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 617
Illegal BamHI site found at 1
Illegal BamHI site found at 1053 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Usage and Biology
Figure 1: Control, non-induced Hwp1 expressing yeast. Dispersed, no major
clustering.
Figure 2: Experiment, induced Hwp1
expressing yeast. 3-D cluster of yeast cells
growing around and on top of other
cells. 2020 XHD-Wuhan-China’s Contribution