Extraction and purification of queen venom and worker venom protein (Sol i II)

Solenopsis Invicta is a serious invasive species which causes anaphylaxis (venom allergic reactions) with its painful sting. In 1988, Hoffman, et al. isolated the four different types of proteins in fire ant venom and noted that these proteins have significant allergenic activity towards immunoglobulin E (Ig E). Among these four proteins, two of them were characterized. This research project focused on expression and purification of one of the uncharacterized proteins, (Sol i II) found in both Worker Venom and Queen Venom variants. My research started with the production of Queen and Worker synthetic Sol i II genes, optimized for expression in E. coli by assembly PCR and then cloned into BL-21 cells for protein expression. However, when the protein was expressed in these cells, it also contains endotoxin in the end product hindering future research work with this protein. To overcome this I am attempting to transform these Sol i II genes into Bacillus megaterium to produce an endotoxin-free final product.