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Team:HKUST/Back4
From 2009.igem.org
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The binary toxin BinA and BinB, which is produced in Bacillus sphaericus, is a mosquito-larvicidal crystal protein. It has its maximum activity when both components are present in equimolar ratio[1]. It could kill larvae by forming pores once binA binds to and binB inserts to the membrane lipid bilayer, leading to swelling and lysis of the cell[2].
Attempts have been made to produce the binary toxin at a high level in many organisms, including Escherichia coli[3]. To enhance expression level and toxin potency, one approach is improving toxin solubility by tagging with T7 tag [4] or glutathione S-transferase (GST), between which GST seems to work better. Not only does GST increase the protein expression level, it improves the solubility of these proteins as well[2].
S. cerevisiae has also been a good tool for bacterial toxin studies, for it can functionally express many bacterial toxins[5]. Since the binary toxin Bin A and Bin B are very specific to insects, we propose that yeast can express this toxin without damaging itself, while insects that eat this yeast will be killed by the toxin. Therefore, we decide to choose this binary toxin as our “insect killer”.
Background
Experimental Design
Parts Design
Future Work
References
Attempts have been made to produce the binary toxin at a high level in many organisms, including Escherichia coli[3]. To enhance expression level and toxin potency, one approach is improving toxin solubility by tagging with T7 tag [4] or glutathione S-transferase (GST), between which GST seems to work better. Not only does GST increase the protein expression level, it improves the solubility of these proteins as well[2].
S. cerevisiae has also been a good tool for bacterial toxin studies, for it can functionally express many bacterial toxins[5]. Since the binary toxin Bin A and Bin B are very specific to insects, we propose that yeast can express this toxin without damaging itself, while insects that eat this yeast will be killed by the toxin. Therefore, we decide to choose this binary toxin as our “insect killer”.
iGEM 2009
