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- | <html>
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- | <head>
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- |
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- | <style>
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| |
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| |
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| |
- |
| |
- | <div class=Section1>
| |
- |
| |
- | <div style='mso-element:para-border-div;border:none;border-bottom:solid #4F81BD 1.0pt;
| |
- | mso-border-bottom-themecolor:accent1;padding:0cm 1cm 4.0pt 0cm'>
| |
- |
| |
- | <embed src="https://static.igem.org/mediawiki/2010/5/51/Nish1.swf" width=900 height=500>
| |
- |
| |
- | <p align=center><img src="https://static.igem.org/mediawiki/2010/5/58/IGEM_IITD_logo.png" width="100"/></p>
| |
- |
| |
- | <p class=MsoTitle align=center style='text-align:center'><span lang=EN-US><i style='mso-bidi-font-style:
| |
- | normal'><b>Dr. Coli </b><o:p></o:p></i></span></p>
| |
- |
| |
- | </div>
| |
- |
| |
- | <p class=MsoNormal><span lang=EN-US><o:p> </o:p></span></p>
| |
- |
| |
- | <h2><span lang=EN-US><u>Renin Angiotensin Aldosterone System (RAAS)<o:p></o:p></span></u></h2>
| |
- |
| |
- | <p class=MsoNormal><span lang=EN-US><o:p> </o:p></span></p>
| |
- |
| |
- | <h3><span lang=EN-US>Introduction</span></h3>
| |
- |
| |
- | <p class=MsoNormal style='text-align:justify'><span lang=EN-US>The Renin
| |
- | Angiotensin Aldosterone System is a hormone system in the human body that is
| |
- | responsible for the regulation of blood pressure and fluid balance.</span></p>
| |
- |
| |
- | <h3 style='text-align:justify'><span lang=EN-US>Functioning</span></h3>
| |
- |
| |
- | <p class=MsoNormal style='text-align:justify'><span lang=EN-US>A decrease in
| |
- | renal perfusion causes the release of the enzyme renin which subsequently
| |
- | cleaves angiotensinogen to angiotensin I. This is further converted to
| |
- | angiotensin II by angiotensin-converting enzyme (ACE). The bioactive product,
| |
- | angiotensin II, causes the constriction of blood vessels, leading to an
| |
- | increased blood pressure. It is also responsible for the secretion of
| |
- | aldosterone by the adrenal cortex, which acts by increasing the re-absorption
| |
- | of sodium and water into the blood. Hence, the fluid volume as well as the
| |
- | blood pressure is maintained in the body.</span></p>
| |
- |
| |
- | <h2><u><span lang=EN-US>The α-hemolysin System in <i style='mso-bidi-font-style:
| |
- | normal'>E.coli<o:p></o:p></i></span></u></h2>
| |
- |
| |
- | <p class=MsoNormal><span lang=EN-US><o:p> </o:p></span></p>
| |
- |
| |
- | <h3><span lang=EN-US>Introduction</span></h3>
| |
- |
| |
- | <p class=MsoNormal style='text-align:justify'><span lang=EN-US>The
| |
- | alpha-hemolysin system is a type I secretion system found in wild type <i
| |
- | style='mso-bidi-font-style:normal'>E.coli </i>which enable them to cause
| |
- | infection in the urinary tract and is an important virulence factor because of
| |
- | its cytotoxic and cytolytic activity against a variety of mammalian cells. </span></p>
| |
- |
| |
- | <h3 style='text-align:justify'><span lang=EN-US>Functioning</span></h3>
| |
- |
| |
- | <p class=MsoNormal style='text-align:justify'><span lang=EN-US>The hemolysin
| |
- | system originates from a series of hly genes. These genes are present in the
| |
- | series <i style='mso-bidi-font-style:normal'>hlyC – hlyA – hlyB – hlyD</i>. Of
| |
- | these, <i style='mso-bidi-font-style:normal'>hlyA</i> codes for the protein
| |
- | that is responsible for the infection. Hly B and D are membrane proteins that
| |
- | work in coordination for the secretion of HlyA. The synthesis, activation and
| |
- | secretion of HlyA in <i style='mso-bidi-font-style:normal'>E.coli </i>is
| |
- | determined by the <i style='mso-bidi-font-style:normal'>hlyCABD</i> operon. This
| |
- | operon is either located either on the chromosome or on transmissible plasmids.
| |
- |
| |
- | </span></p>
| |
- |
| |
- | <p class=MsoNormal style='text-align:justify'><span lang=EN-US>It has been
| |
- | shown that this system can be used to produce and secrete a protein of our
| |
- | choice if the protein contains 60 amino acid residues from the carboxy terminal
| |
- | end of the HlyA protein. </span></p>
| |
- |
| |
- | <h2><u><span lang=EN-US>Our iGEM 2010 Project<o:p></o:p></span></u></h2>
| |
- |
| |
- | <p class=MsoNormal><span lang=EN-US><o:p> </o:p></span></p>
| |
- |
| |
- | <h3><span lang=EN-US>Introduction</span></h3>
| |
- |
| |
- | <p class=MsoNormal style='text-align:justify'><span lang=EN-US>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. </span></p>
| |
- |
| |
- | <h3><span lang=EN-US>Approach</span></h3>
| |
- |
| |
- | <p class=MsoNormal><span lang=EN-US>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.</span></p>
| |
- |
| |
- | <p class=MsoNormal><span lang=EN-US>For this we will be introducing two vectors
| |
- | into <i style='mso-bidi-font-style:normal'>E. coli </i>as shown below.</span></p>
| |
- |
| |
- | <p class=MsoNormal><span style='mso-ansi-language:EN-IN;mso-fareast-language:
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| |
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| |
- | lang=EN-US><span style='mso-spacerun:yes'> </span><span
| |
- | style='mso-tab-count:1'> </span></span><span style='mso-ansi-language:
| |
- | EN-IN;mso-fareast-language:EN-IN;mso-bidi-language:HI;mso-no-proof:yes'><!--[if gte vml 1]><v:shape
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| |
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| |
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| |
- |
| |
- | <p class=MsoNormal><span lang=EN-US><o:p> </o:p></span></p>
| |
- |
| |
- | <p class=MsoNormal style='text-align:justify'><span lang=EN-US>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.</span></p>
| |
- |
| |
- | <p class=MsoNormal><span lang=EN-US><o:p> </o:p></span></p>
| |
- |
| |
- | <p class=MsoNormal><span lang=EN-US>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.</span></p>
| |
- |
| |
- | <p class=MsoNormal><span lang=EN-US>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</span></p>
| |
- |
| |
- | <p class=MsoNormal><span lang=EN-US>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.</span></p>
| |
- |
| |
- | <p class=MsoNormal><span lang=EN-US><o:p> </o:p></span></p>
| |
- |
| |
- | <p class=MsoNormal style='text-align:justify'><span lang=EN-US><o:p> </o:p></span></p>
| |
- |
| |
- | </div>
| |
- |
| |
- | </body>
| |
- |
| |
- | </html>
| |
- |
| |
- |
| |
- | {| style="color:#1b2c8a;background-color:#0c6;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"
| |
- | !align="center"|[[Team:IIT_Delhi_1|Home]]
| |
- | !align="center"|[[Team:IIT_Delhi_1/Team|Team]]
| |
- | !align="center"|[https://igem.org/Team.cgi?year=2010&team_name=IIT_Delhi_1 Official Team Profile]
| |
- | !align="center"|[[Team:IIT_Delhi_1/Project|Project]]
| |
- | !align="center"|[[Team:IIT_Delhi_1/Parts|Parts Submitted to the Registry]]
| |
- | !align="center"|[[Team:IIT_Delhi_1/Modeling|Modeling]]
| |
- | !align="center"|[[Team:IIT_Delhi_1/Notebook|Notebook]]
| |
- | !align="center"|[[Team:IIT_Delhi_1/Safety|Safety]]
| |
- | |}
| |
- | ''Italic text''
| |