Horseshoe Crab Factor-C Detection

First, we will synthetically produce the factor C protein (from the horseshoe crab) using overlap extension PCR. A fusion of factor c and hCG beta subunit will be synthesized and used as the detection system for gram-negative bacteria in tandem with pregnancy tests.

Bacteria vs. Tobacco, Mambalgin-1 Production Showdown!

We will engineer two different chassis systems to produce a eukaryotic analgesic protein derived from Dendroaspis polylepis. The first will consist of transformed Escherichia coli that will assemble our protein and localize it to the periplasm. This method should allow for more efficient disulfide bond formation due to the periplasm's oxidizing environment. Furthermore, this strategy will reduce the labor required to isolate our protein during purification. Meanwhile, the second system will consist of Agrobacterium transfected Nicotiana benthamiana plants. The infiltrated leaves will produce our protein of interest over several days. Then, the leaves will be collected, macerated and further purified to isolate the protein product. Finally, by using two systems that differ by domain, we can determine which organism-based chassis is superior for producing our protein of interest.