Project Description

The goal of our project is to create an easy, cost-effective, and sensitive contamination detection device for iGEM teams using Human Chorionic Gonadotropin (HCG). After looking into the properties of Horseshoe crab blood, we found that lipopolysaccharides (LPS) activate a protein known as Factor C. Factor C is sensitive to the LPS; in the presence of LPS Factor C undergoes auto-catalysis which initiates the cascade of protein activation steps leading to coagulation. To replace this complicated cascade, we propose to devise a detection system that signals auto-catalysis of Factor C. We propose to synthesize a fusion protein of Factor C and HCG which when activated by auto-catalysis will give a positive signal on a pregnancy test strip. We will first test various chains of HCG at different concentrations and see how they vary in sensitivity to the pregnancy test "Clinical Guard." Then based on our results, we will pair the most effective chain of HCG with Factor C. Then we will insert the DNA sequence into pSB1C3 for submission, and pGEX for protein expression. From there we will purify the protein produced by the pGEX. To produce a detection system we will fuse the purified protein to the membrane strip and insert it into a saline solution. Then to check for contamination, a pregnancy test strip will be inserted into the solution and will trigger a response based upon the presence of LPS. .

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