Difference between revisions of "Team:Georgia State"

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<h2>Project Description</h2>
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<h2>Is Your Detector Expecting? See with HCG!</h2>
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<p>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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Detection is used in all aspects of biology and is essential in providing an illustration of the chemical world around us. Currently, fluorescent protein are used as reporters but they require additional analysis with expensive and immobile equipment. We propose to create an alternative detection system kit using recombinant Human Chorionic Gonadotropin (HCG) as a reporter. The goal of our project is to create an easy, cost-effective, and sensitive detection device for use in synthetic biology, it can even be used by other iGEM teams to get an all-or-nothing response to indicate the presence or absence of targeted protein. We plan to create a pGEX plasmid containing the recombinant beta subunit of HCG with a strong promoter as a positive promoter and an additional plasmid containing recombinant HCG preceded by restriction sites not present elsewhere in the plasmid and this will be where the promoter is inserted. The promoter will only synthesize HCG when it is activated by the presence of the protein in question. Then when a pregnancy test strip is inserted in the sample, it will trigger the response based on the activation of the introduced promoter.                                                
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<p><a href="https://www.youtube.com/watch?v=ntA4nKMk93Q" class="btn btn-special popup-vimeo">Watch The Video</a></p>
 
<p><a href="https://www.youtube.com/watch?v=ntA4nKMk93Q" class="btn btn-special popup-vimeo">Watch The Video</a></p>

Revision as of 04:10, 15 September 2018

Is Your Detector Expecting? See with HCG!

Detection is used in all aspects of biology and is essential in providing an illustration of the chemical world around us. Currently, fluorescent protein are used as reporters but they require additional analysis with expensive and immobile equipment. We propose to create an alternative detection system kit using recombinant Human Chorionic Gonadotropin (HCG) as a reporter. The goal of our project is to create an easy, cost-effective, and sensitive detection device for use in synthetic biology, it can even be used by other iGEM teams to get an all-or-nothing response to indicate the presence or absence of targeted protein. We plan to create a pGEX plasmid containing the recombinant beta subunit of HCG with a strong promoter as a positive promoter and an additional plasmid containing recombinant HCG preceded by restriction sites not present elsewhere in the plasmid and this will be where the promoter is inserted. The promoter will only synthesize HCG when it is activated by the presence of the protein in question. Then when a pregnancy test strip is inserted in the sample, it will trigger the response based on the activation of the introduced promoter. .

Watch The Video

Marketing

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