Team:IIT Delhi 1

Revision as of 18:53, 26 October 2010 (view source)

Nishantbpl (Talk | contribs)

← Older edit

Revision as of 00:56, 28 October 2010 (view source)

Nishantbpl (Talk | contribs)

Newer edit →

Line 557:

mso-border-bottom-themecolor:accent1;padding:0cm 1cm 4.0pt 0cm'>

-

+

-

~~<i style='mso-bidi-font-style:~~

-

~~normal'>Dr. Coli ~~

-

~~</div>~~

-

~~~~

-

~~<h2>Renin Angiotensin Aldosterone System (RAAS)<o:p></o:p></h2>~~

-

~~<o:p> </o:p>~~

-

~~<h3>Introduction</h3>~~

-

~~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.~~

-

~~<h3 style='text-align:justify'>Functioning</h3>~~

-

~~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.~~

-

~~<h2>The α-hemolysin System in <i style='mso-bidi-font-style:~~

-

~~normal'>E.coli<o:p></o:p></h2>~~

-

~~<o:p> </o:p>~~

-

~~<h3>Introduction</h3>~~

-

~~The~~

-

~~alpha-hemolysin system is a type I secretion system found in wild type <i~~

-

~~style='mso-bidi-font-style:normal'>E.coli 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. ~~

-

~~<h3 style='text-align:justify'>Functioning</h3>~~

-

~~The hemolysin~~

-

~~system originates from a series of hly genes. These genes are present in the~~

-

~~series hlyC – hlyA – hlyB – hlyD. Of~~

-

~~these, hlyA 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 E.coli is~~

-

~~determined by the hlyCABD operon. This~~

-

~~operon is either located either on the chromosome or on transmissible plasmids.~~

-

~~~~

-

~~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. ~~

-

~~<h2>Our iGEM 2010 Project<o:p></o:p></h2>~~

-

~~<o:p> </o:p>~~

-

~~<h3>Introduction</h3>~~

-

~~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. ~~

-

~~<h3>Approach</h3>~~

-

~~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 below.~~

-

~~<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:shapetype~~

-

~~id="_x0000_t75" coordsize="21600,21600" o:spt="75" o:preferrelative="t"~~

-

~~path="m@4@5l@4@11@9@11@9@5xe" filled="f" stroked="f">~~

-

~~<v:stroke joinstyle="miter"/>~~

-

~~<v:formulas>~~

-

~~<v:f eqn="if lineDrawn pixelLineWidth 0"/>~~

-

~~<v:f eqn="sum @0 1 0"/>~~

-

~~<v:f eqn="sum 0 0 @1"/>~~

-

~~<v:f eqn="prod @2 1 2"/>~~

-

~~<v:f eqn="prod @3 21600 pixelWidth"/>~~

-

~~<v:f eqn="prod @3 21600 pixelHeight"/>~~

-

~~<v:f eqn="sum @0 0 1"/>~~

-

~~<v:f eqn="prod @6 1 2"/>~~

-

~~<v:f eqn="prod @7 21600 pixelWidth"/>~~

-

~~<v:f eqn="sum @8 21600 0"/>~~

-

~~<v:f eqn="prod @7 21600 pixelHeight"/>~~

-

~~<v:f eqn="sum @10 21600 0"/>~~

-

~~</v:formulas>~~

-

~~<v:path o:extrusionok="f" gradientshapeok="t" o:connecttype="rect"/>~~

-

~~<o:lock v:ext="edit" aspectratio="t"/>~~

-

~~</v:shapetype><v:shape id="Picture_x0020_0" o:spid="_x0000_i1026" type="#_x0000_t75"~~

-

~~alt="Vector1.png" style='width:195pt;height:209.25pt;visibility:visible;~~

-

~~mso-wrap-style:square'>~~

-

~~<v:imagedata src="IIT+Delhi+iGEM+Wiki_files/image001.png" o:title="Vector1"/>~~

-

~~</v:shape><![endif]--><![if !vml]><img width=260 height=279~~

-

~~src="IIT+Delhi+iGEM+Wiki_files/image002.gif" alt=Vector1.png v:shapes="Picture_x0020_0"><![endif]><span~~

-

~~lang=EN-US> <span~~

-

~~style='mso-tab-count:1'> <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~~

-

~~id="Picture_x0020_1" o:spid="_x0000_i1025" type="#_x0000_t75" alt="Vector2.png"~~

-

~~style='width:195pt;height:212.25pt;visibility:visible;mso-wrap-style:square'>~~

-

~~<v:imagedata src="IIT+Delhi+iGEM+Wiki_files/image003.png" o:title="Vector2"/>~~

-

~~</v:shape><![endif]--><![if !vml]><img width=260 height=283~~

-

~~src="IIT+Delhi+iGEM+Wiki_files/image004.gif" alt=Vector2.png v:shapes="Picture_x0020_1"><![endif]>~~

-

~~<o:p> </o:p>~~

-

~~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.~~

-

~~<o:p> </o:p>~~

-

~~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.~~

-

~~<o:p> </o:p>~~

-

~~<o:p> </o:p>~~

</div>

Line 717:

Line 565:

</html>

-

~~''Italic text''~~

@@ Line 557: / Line 557: @@
 mso-border-bottom-themecolor:accent1;padding:0cm 1cm 4.0pt 0cm'>
-<p align=center><embed src="https://static.igem.org/mediawiki/2010/5/51/Nish1.swf" width=900 height=500></p>
+<p align=center><embed src="https://static.igem.org/mediawiki/igem.org/5/51/Nish1.swf" width=900 height=500></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></i></span></p>
-</div>
-<p class=MsoNormal><span lang=EN-US></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>&nbsp;</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 &#945;-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>&nbsp;</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>&nbsp;</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:
-EN-IN;mso-bidi-language:HI;mso-no-proof:yes'><!--[if gte vml 1]><v:shapetype
- id="_x0000_t75" coordsize="21600,21600" o:spt="75" o:preferrelative="t"
- path="m@4@5l@4@11@9@11@9@5xe" filled="f" stroked="f">
- <v:stroke joinstyle="miter"/>
- <v:formulas>
-  <v:f eqn="if lineDrawn pixelLineWidth 0"/>
-  <v:f eqn="sum @0 1 0"/>
-  <v:f eqn="sum 0 0 @1"/>
-  <v:f eqn="prod @2 1 2"/>
-  <v:f eqn="prod @3 21600 pixelWidth"/>
-  <v:f eqn="prod @3 21600 pixelHeight"/>
-  <v:f eqn="sum @0 0 1"/>
-  <v:f eqn="prod @6 1 2"/>
-  <v:f eqn="prod @7 21600 pixelWidth"/>
-  <v:f eqn="sum @8 21600 0"/>
-  <v:f eqn="prod @7 21600 pixelHeight"/>
-  <v:f eqn="sum @10 21600 0"/>
- </v:formulas>
- <v:path o:extrusionok="f" gradientshapeok="t" o:connecttype="rect"/>
- <o:lock v:ext="edit" aspectratio="t"/>
-</v:shapetype><v:shape id="Picture_x0020_0" o:spid="_x0000_i1026" type="#_x0000_t75"
- alt="Vector1.png" style='width:195pt;height:209.25pt;visibility:visible;
- mso-wrap-style:square'>
- <v:imagedata src="IIT+Delhi+iGEM+Wiki_files/image001.png" o:title="Vector1"/>
-</v:shape><![endif]--><![if !vml]><img width=260 height=279
-src="IIT+Delhi+iGEM+Wiki_files/image002.gif" alt=Vector1.png v:shapes="Picture_x0020_0"><![endif]></span><span
-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
- id="Picture_x0020_1" o:spid="_x0000_i1025" type="#_x0000_t75" alt="Vector2.png"
- style='width:195pt;height:212.25pt;visibility:visible;mso-wrap-style:square'>
- <v:imagedata src="IIT+Delhi+iGEM+Wiki_files/image003.png" o:title="Vector2"/>
-</v:shape><![endif]--><![if !vml]><img width=260 height=283
-src="IIT+Delhi+iGEM+Wiki_files/image004.gif" alt=Vector2.png v:shapes="Picture_x0020_1"><![endif]></span></p>
-<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</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>&nbsp;</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>&nbsp;</o:p></span></p>
-<p class=MsoNormal style='text-align:justify'><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
 </div>
@@ Line 717: / Line 565: @@
 </html>
-''Italic text''

From 2010.igem.org

Revision as of 00:56, 28 October 2010