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Team:Osaka/SIGNAL
From 2009.igem.org
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<h1 style="text-align: left">SIGNAL</h1> | <h1 style="text-align: left">SIGNAL</h1> | ||
| - | <span> | + | <span>We will be using signaling parts from quorum sensing systems of various bacteria to implement intracellular communication between bacterial colonies of different 'colors'. If it works, we can for example cause two colonies of bacteria to change color or stop moving as they approach each other, hopefully resulting in interesting patterns.<br><br> |
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| + | Currently we are working on 2 distinct groups of parts: 'Senders' and 'Receivers'. 'Senders' code for enzymes that produce AHL signal molecules, which diffuse out of the cell, through the culture medium and into the receiving cell, where a receptor protein encoded by the 'Receiver' binds the signals, forming a complex which in turn can bind to and upregulate transcription from specific promoter.<br><br> | ||
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| + | To test the effectiveness of our signaling system, we will first test the Receivers by attaching a GFP coding unit (rbs + protein coding region + terminator) behind the Receiver's AHL-activated promoter. We will then directly add chemically-derived AHL into culture solutions of these 'Test Receivers' and look for GFP fluorescence which will indicate that transcription has been activated by AHL.<br><br> | ||
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| + | Following that, we will test the Senders by using the above Receivers. We can determine the amount of AHL produced by the Senders by comparing transcription activity induced by the Senders in relation to AHL-induced trancription activity.<br><br> | ||
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| + | Once both Sender and Receiver function has been confirmed and characterized, we will then attempt to characterize the function of the whole system in a way that relates to our intended usage. We will spot two colonies, one of Senders and one of Receivers, on a soft agar plate and determine the maximum distance that the Senders can successfully activate the Receivers. Of course, this will only work if the AHL molecules can effectively diffuse through the agar medium.<br><br> | ||
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| + | Stay tuned for more updates! :)</span> | ||
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Revision as of 10:11, 30 September 2009
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SIGNAL
We will be using signaling parts from quorum sensing systems of various bacteria to implement intracellular communication between bacterial colonies of different 'colors'. If it works, we can for example cause two colonies of bacteria to change color or stop moving as they approach each other, hopefully resulting in interesting patterns.Currently we are working on 2 distinct groups of parts: 'Senders' and 'Receivers'. 'Senders' code for enzymes that produce AHL signal molecules, which diffuse out of the cell, through the culture medium and into the receiving cell, where a receptor protein encoded by the 'Receiver' binds the signals, forming a complex which in turn can bind to and upregulate transcription from specific promoter.
To test the effectiveness of our signaling system, we will first test the Receivers by attaching a GFP coding unit (rbs + protein coding region + terminator) behind the Receiver's AHL-activated promoter. We will then directly add chemically-derived AHL into culture solutions of these 'Test Receivers' and look for GFP fluorescence which will indicate that transcription has been activated by AHL.
Following that, we will test the Senders by using the above Receivers. We can determine the amount of AHL produced by the Senders by comparing transcription activity induced by the Senders in relation to AHL-induced trancription activity.
Once both Sender and Receiver function has been confirmed and characterized, we will then attempt to characterize the function of the whole system in a way that relates to our intended usage. We will spot two colonies, one of Senders and one of Receivers, on a soft agar plate and determine the maximum distance that the Senders can successfully activate the Receivers. Of course, this will only work if the AHL molecules can effectively diffuse through the agar medium.
Stay tuned for more updates! :)
