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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 yeast and induced by galactose. We will use western blotting to test toxin expressing with an 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 toxin-expressing yeast cells 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].

  • Background
  • Experimental Design
  • Parts Design
  • Future Work
  • References