Difference between revisions of "Team:NAWI Graz/Design"

Line 1: Line 1:
 
{{NAWI_Graz:navbar}}
 
{{NAWI_Graz:navbar}}
 
<html>
 
<html>
 +
 +
    <div class="section section-heading container">
 +
        <h1>DESIGN</h1>
 +
    </div>
 +
    <br>
 +
 +
    <div class="section container">
 +
        <div class="section-text container">
 +
            <p>The bacteria-robot interface is realized as a highly modular feedback system.</p>
 +
            <p>A mobile robot’s sensor values alter the environment of ‘’’E. coli’’’ strains that were designed to respond to
 +
                these changes with increased expression of fluorescent proteins. In turn, this fluorescence is measured and
 +
                its quantities are translated into robot behaviour.</p>
 +
            <p>For how it all played out, see [[Results]], but let’s first have a glimpse at the modules comprising the feedback
 +
                loop (for a full description see [[Parts]]):</p>
 +
        </div>
 +
    </div>
 +
    <br>
 +
 +
    <div class="section container">
 +
        <h2 class="section-sub">The Bacteria</h2>
 +
        <div class="section-text container">
 +
            <div class="section-link">([[Parts#Bacteria|‘’full description’’]])</div>
 +
            Our cultures have one goal: they should react to changes in their environment by expressing fluorescent proteins.  Therefore,
 +
            we introduced three promoters, two sensing the pH and one sensing the temperature of the culture media. Activation
 +
            of one promoter leads to transcription and translation of a fluorescence protein.
 +
            <h3 class="section-sub-sub">"ibpA" - Heat Shock Promoter</h3>
 +
            <div class="section-sub-text container">
 +
                The heat shock promoter ibpA is controlled by the transcription factor σ32. In principle, the exposure to high temperatures
 +
                leads to an increase of σ32, which subsequently enables heat shock promoters to be recognized by the RNA
 +
                polymerase. The promoter exhibits a high induction rate and high levels of expression. In our experiment,
 +
                the ibpA promoter controls the expression of GFP.
 +
            </div>
 +
            <h3 class="section-sub-sub">"asr" - Acid Inductible Promoter</h3>
 +
            <div class="section-sub-text container">
 +
                <div class="row">
 +
                    <div class="col">
 +
                        Promoter activity is controlled by the RstAB System detecting the pH and the PhoRB System activated when inorganic phosphate
 +
                        is rare. Thus, expression only works in low phosphate media (LPM). When grown in LPM and activated
 +
                        by a switch of pH to 5,5 the promoter becomes active and mCardinal is expressed. To enhance expression
 +
                        an extra ribosome binding site (RBS) was inserted between the promoter and mCardinal.
 +
                    </div>
 +
                    <div class="col">
 +
                        <img src="" alt="[asr inductible promoter]">
 +
                    </div>
 +
                </div>
 +
            </div>
 +
            <h3 class="section-sub-sub">"alx" - Alkaline-induced Roboswitch</h3>
 +
            <div class="section-sub-text container">
 +
                <div class="row">
 +
                    <div class="col">
 +
                        <img src="" alt="[alx riboswitch]">
 +
                    </div>
 +
                    <div class="col">
 +
                        Promoter activity is controlled by the RstAB System detecting the pH and the PhoRB System activated when inorganic phosphate
 +
                        is rare. Thus, expression only works in low phosphate media (LPM). When grown in LPM and activated
 +
                        by a switch of pH to 5,5 the promoter becomes active and mCardinal is expressed. To enhance expression
 +
                        an extra ribosome binding site (RBS) was inserted between the promoter and mCardinal.
 +
                    </div>
 +
                </div>
 +
            </div>
 +
        </div>
 +
    </div>
 +
    <br>
 +
 +
    <div class="section container">
 +
        <h2 class="section-sub">The Bioreactor</h2>
 +
        <div class="section-text container">
 +
            <div class="section-link">([[Parts#Bioreactor|‘’full description’’]])</div>
 +
            <h3 class="section-sub-sub">subsubsection</h3>
 +
            <div class="section-sub-text container">
 +
 +
            </div>
 +
            <h3 class="section-sub-sub">"subsubsection</h3>
 +
            <div class="section-sub-text container">
 +
                <div class="row">
 +
                    <div class="col">
 +
                        texttexttext
 +
                    </div>
 +
                    <div class="col">
 +
                        <img src="" alt="[image]">
 +
                    </div>
 +
                </div>
 +
            </div>
 +
        </div>
 +
    </div>
 +
    <br>
 +
 +
    <div class="section container">
 +
        <h2 class="section-sub">TITLE</h2>
 +
        <div class="section-text container">
 +
 +
        </div>
 +
    </div>
 +
 
 +
 +
 +
 +
  
  

Revision as of 11:13, 28 October 2017

DESIGN


The bacteria-robot interface is realized as a highly modular feedback system.

A mobile robot’s sensor values alter the environment of ‘’’E. coli’’’ strains that were designed to respond to these changes with increased expression of fluorescent proteins. In turn, this fluorescence is measured and its quantities are translated into robot behaviour.

For how it all played out, see [[Results]], but let’s first have a glimpse at the modules comprising the feedback loop (for a full description see [[Parts]]):


The Bacteria

([[Parts#Bacteria|‘’full description’’]])
Our cultures have one goal: they should react to changes in their environment by expressing fluorescent proteins.  Therefore, we introduced three promoters, two sensing the pH and one sensing the temperature of the culture media. Activation of one promoter leads to transcription and translation of a fluorescence protein.

"ibpA" - Heat Shock Promoter

The heat shock promoter ibpA is controlled by the transcription factor σ32. In principle, the exposure to high temperatures leads to an increase of σ32, which subsequently enables heat shock promoters to be recognized by the RNA polymerase. The promoter exhibits a high induction rate and high levels of expression. In our experiment, the ibpA promoter controls the expression of GFP.

"asr" - Acid Inductible Promoter

Promoter activity is controlled by the RstAB System detecting the pH and the PhoRB System activated when inorganic phosphate is rare. Thus, expression only works in low phosphate media (LPM). When grown in LPM and activated by a switch of pH to 5,5 the promoter becomes active and mCardinal is expressed. To enhance expression an extra ribosome binding site (RBS) was inserted between the promoter and mCardinal.
[asr inductible promoter]

"alx" - Alkaline-induced Roboswitch

[alx riboswitch]
Promoter activity is controlled by the RstAB System detecting the pH and the PhoRB System activated when inorganic phosphate is rare. Thus, expression only works in low phosphate media (LPM). When grown in LPM and activated by a switch of pH to 5,5 the promoter becomes active and mCardinal is expressed. To enhance expression an extra ribosome binding site (RBS) was inserted between the promoter and mCardinal.

The Bioreactor

([[Parts#Bioreactor|‘’full description’’]])

subsubsection

"subsubsection

texttexttext
[image]

TITLE

Design

Design is the first step in the design-build-test cycle in engineering and synthetic biology. Use this page to describe the process that you used in the design of your parts. You should clearly explain the engineering principles used to design your project.

This page is different to the "Applied Design Award" page. Please see the Applied Design page for more information on how to compete for that award.

What should this page contain?
  • Explanation of the engineering principles your team used in your design
  • Discussion of the design iterations your team went through
  • Experimental plan to test your designs
Inspiration