Composite part
We have known the function of the three toxic peptides of Pantide, ω-hexatoxin-Hv1a, µ-segestritoxin-Sf1a, and Orally Active Insecticidal Peptide (OAIP) respectively and they are easy to express with the help of the bacteria E. coli BL21 Rosetta Gami strain. So, we then start to create BioBricks of these three toxins which we have obtained the nucleotide sequences. We use a strong T7 promoter that is suitable with the strain of E. coli we chose and also powerful at expressing proteins with multiple disulfide bonds, and we add His-tag for protein purification.
Pantide without lectin
Pantide can be a single toxic peptide of the three toxins we chose, Hv1a, Sf1a, and OAIP respectively, which derive from the natural predators spiders.
BBa_........
T7 promoter+RBS+Hv1a+linker+His-tag+terminator
Pantide that contains the gene coding for Hv1a linked with His-tag.
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T7 promoter+RBS+Sf1a+linker+His-tag+terminator
Pantide that contains the gene coding for Sf1a linked with His-tag.
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T7 promoter+RBS+OAIP+linker+His-tag+terminator
Pantide that contains the gene coding for OAIP linked with His-tag.
Pantide with lectin
Pantide can be a single toxic peptide of the three toxins we chose, Hv1a, Sf1a, and OAIP respectively with the addition of Galanthus nivalis agglutinin (GNA), for GNA has the function of acting as a carrier of Pantide from insect gut to the haemolymph and finally Pantide can transmit to the action site.
BBa_........
T7 promoter+RBS+Hv1a-lectin+linker+His-tag+terminator
Pantide that contains the gene coding for Hv1a and the snowdrop lectin linked with His-tag.
BBa_........
T7 promoter+RBS+Sf1a-lectin+linker+His-tag+terminator
Pantide that contains the gene coding for Sf1a and the snowdrop lectin linked with His-tag.
BBa_........
T7 promoter+RBS+OAIP-lectin+linker+His-tag+terminator
Pantide that contains the gene coding for OAIP and the snowdrop lectin linked with His-tag.
Pantide with Gs linker
In this part, to facilitate the expression of Pantide, we extended the linker with the substitution of Gs linker that is even better than the original linker for the more correcter folding of the proteins structure.
BBa_........
T7 promoter+RBS+Hv1a-lectin+Gs linker+His-tag+terminator
Pantide that contains the gene coding for Hv1a and the snowdrop lectin. And improve the correct folding of Pantide by substituting with Gs linker linked with His-tag.
