"
Team:HKUST/Group4
From 2009.igem.org
(Difference between revisions)
| Line 56: | Line 56: | ||
<p>Protein Expression</p> | <p>Protein Expression</p> | ||
We have designed several constructs to test the binary protein's toxicity to insects as well as to yeast:<br> | We have designed several constructs to test the binary protein's toxicity to insects as well as to yeast:<br> | ||
| - | |||
| - | |||
| - | + | ||
| - | <img src="http://igem2009hkust.fileave.com/wiki/Group4/ | + | <img src="http://igem2009hkust.fileave.com/wiki/Group4/figure01.jpg " width=588; height=400 /></a><br> |
| - | + | ||
| - | + | ||
These constructs will be transformed into the yeast and induced by galactose. Then we will use western blot to test the expression of toxin by anti-GST antibody. We can estimate the protein level as well as the yeast phenotype: whether they are resistant to the toxin.</p> | These constructs will be transformed into the yeast and induced by galactose. Then we will use western blot to test the expression of toxin by anti-GST antibody. We can estimate the protein level as well as the yeast phenotype: whether they are resistant to the toxin.</p> | ||
<br> | <br> | ||
<p>Drosophila Larvicidal Essay</p> | <p>Drosophila Larvicidal Essay</p> | ||
Next we will test the biological function of toxin produced in yeast to kill Drosophila larvae. The Drosophila larvicidal activity is tested by diluting yeast cells expressing the toxin in water and adding them to water containing 10 larvae in each well of a 24-well tissue culture plate (diameter of the well is 1.5 cm)(Figure 1). Mortality is recorded after incubation at 30°C for 48 h. LC50 is determined by Probit analysis[6].</p> | Next we will test the biological function of toxin produced in yeast to kill Drosophila larvae. The Drosophila larvicidal activity is tested by diluting yeast cells expressing the toxin in water and adding them to water containing 10 larvae in each well of a 24-well tissue culture plate (diameter of the well is 1.5 cm)(Figure 1). Mortality is recorded after incubation at 30°C for 48 h. LC50 is determined by Probit analysis[6].</p> | ||
| - | <img src="http://igem2009hkust.fileave.com/wiki/Group4/ | + | |
| - | + | <img src="http://igem2009hkust.fileave.com/wiki/Group4/figure02.jpg" width=585; height=505 /></a><br> | |
| + | |||
<li><a href="https://2009.igem.org/Team:HKUST/Back4">Background</a></li> | <li><a href="https://2009.igem.org/Team:HKUST/Back4">Background</a></li> | ||
<li><a href="https://2009.igem.org/Team:HKUST/Group4">Experimental Design</a></li> | <li><a href="https://2009.igem.org/Team:HKUST/Group4">Experimental Design</a></li> | ||
Revision as of 15:24, 21 October 2009
a
Protein Expression
We have designed several constructs to test the binary protein's toxicity to insects as well as to yeast:
These constructs will be transformed into the yeast and induced by galactose. Then we will use western blot to test the expression of toxin by anti-GST antibody. We can estimate the protein level as well as the yeast phenotype: whether they are resistant to the toxin.
Drosophila Larvicidal Essay
Next we will test the biological function of toxin produced in yeast to kill Drosophila larvae. The Drosophila larvicidal activity is tested by diluting yeast cells expressing the toxin in water and adding them to water containing 10 larvae in each well of a 24-well tissue culture plate (diameter of the well is 1.5 cm)(Figure 1). Mortality is recorded after incubation at 30°C for 48 h. LC50 is determined by Probit analysis[6].
iGEM 2009
