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Team:IIT Delhi 1/Project
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Our iGEM 2010 Project
Introduction Our aim is to create an online elicitor detection and response in a flow system, which can function in the detection of high blood pressure and help reduce it by releasing renin inhibitors into the circulatory system. This kind of a system would be helpful to patients with a hypertensive condition.
Approach Since the blood pressure system is not really a chemical signal, what we have assumed is that the blood pressure can be easily converted into a chemical signal. Having assumed this, we will use a chemical signal such as IPTG or AHL as the elicitor to produce appropriate amounts of renin inhibitor. For easy detection and verification, we are planning to use GFP in place of renin inhibitor which can easily be replaced later. For this we will be introducing two vectors into E. coli as shown in the images.
The first vector which contains the GFP-hlyA chimeric sequence with pLac promoter is used to produce the chimera GFP protein which contains 62 amino acid residues from the C terminal of hlyA protein, a prerequisite for the secretion of the protein outside the cell body. Forming a chimera will not in any way change the structure or function as per literature.
The other vector contains the hlyB and hlyD sequences with ptet promoter sequence. These two proteins will be constitutively expressed. These proteins are present in the membrane and provide for an outlet of hlyA or chimeras with 62 amino acid residues of the C terminal of hlyA. After we are successful in obtaining GFP-hlyA chimera outside the cell body, our next task will be to immobilize these cells in a flow system and characterize the flow-expression profile. We would want to keep the cells in G0 phase so as to have continuous expression of the protein without cell death We will also be working towards minimizing the toxins and other products so that such a system can finally be thought fit for incorporating and integrating with the circulatory system in our body.
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