IGEM:Imperial/2010/Experiments1: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
Anita Nguyen (talk | contribs) New page: =Experiments for the Output Amplification Model= * Determine maximum concentration of sD that cells can produce – crucial! (''in vivo''): Compare activities with the next one * Determine... |
Anita Nguyen (talk | contribs) mNo edit summary |
||
| (One intermediate revision by the same user not shown) | |||
| Line 1: | Line 1: | ||
=Experiments for the Output Amplification Model= | <html> | ||
<body style="background-color:FFFFCC"> | |||
Compare | <h1>Experiments for the Output Amplification Model</h1> | ||
In vivo IPTG | <img src="http://upload.wikimedia.org/wikipedia/commons/thumb/4/4f/Postit_large.jpg/500px-Postit_large.jpg"><div style="position:relative;top:-450px;left:50px;font: 11pt helvetica, georgia, courier, arial; margin-left: 10px;"><b>Output Amplification Model</b><br /><br /> | ||
<ol> | |||
<li>Determine the maximum concentration of sD that <br />cells can produce (in vivo): | |||
TEV | Compare activity with 2. | ||
</li> | |||
<li>Determine how much XylE we need to produce <br />prior to adding Catechol (in vitro first, maybe in vivo <br />afterwards). In vivo IPTG: Compare cultures that <br />have less of it. | |||
</li> | |||
<li>Determine XylE and TEV production (in vivo). <br />Use robot to induce production and measure <br />activities TEV (FRET pairs with TEV link). | |||
</li> | |||
<li>Degradation of XylE (in vivo or in vitro if cell division <br />is not taken into account). Monitoring activity, then <br />approximate the concentration. Remove the IPTG. | |||
</li> | |||
<li>Determine TEV kinetics (k<sub>cat</sub>, K<sub>m</sub>) on <br />XylE-GFP (in vitro).</li> | |||
</ol> | |||
</div> | |||
</body> | |||
</html> | |||
Latest revision as of 14:27, 9 September 2010
<html> <body style="background-color:FFFFCC">
Experiments for the Output Amplification Model
<img src="http://upload.wikimedia.org/wikipedia/commons/thumb/4/4f/Postit_large.jpg/500px-Postit_large.jpg">
Output Amplification Model
- Determine the maximum concentration of sD that
cells can produce (in vivo): Compare activity with 2. - Determine how much XylE we need to produce
prior to adding Catechol (in vitro first, maybe in vivo
afterwards). In vivo IPTG: Compare cultures that
have less of it. - Determine XylE and TEV production (in vivo).
Use robot to induce production and measure
activities TEV (FRET pairs with TEV link). - Degradation of XylE (in vivo or in vitro if cell division
is not taken into account). Monitoring activity, then
approximate the concentration. Remove the IPTG. - Determine TEV kinetics (kcat, Km) on
XylE-GFP (in vitro).
</body> </html>
