Team:Bologna
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- | The aim of our project is the design of a new device to control the synthesis of any protein of interest. This "general-purpose" standard device, implemented in <i>E. coli</i>, acts at the translational level to allow a switch in protein expression faster than transcriptional promoter regulation. We named this device <b>T- | + | The aim of our project is the design of a new device to control the synthesis of any protein of interest. This "general-purpose" standard device, implemented in <i>E. coli</i>, acts at the translational level to allow a switch in protein expression faster than transcriptional promoter regulation. We named this device <b>T-REX</b> (<b>T</b>rans <b>R</b>epressor of <b>EX</b>pression). |
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- | '''How T- | + | '''How T-REX works''' |
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Transcription of the target gene yields a mRNA strand - containing the CIS-repressing sequence at its 5' end - available for translation into protein by ribosomes (<i>see Fig. 1, right panel</i>). Induction of the promoter controlling the TRANS coding sequence, releases a transcript complementary to the CIS mRNA sequence. The TRANS/CIS <b>RNA duplex</b> prevents ribosomes from binding to RBS on target mRNA, thus <b>repressing protein synthesis</b>. The amount of the TRANS-repressor regulates the rate of translation of the target mRNA (<i>see Fig. 1, left panel</i>) | Transcription of the target gene yields a mRNA strand - containing the CIS-repressing sequence at its 5' end - available for translation into protein by ribosomes (<i>see Fig. 1, right panel</i>). Induction of the promoter controlling the TRANS coding sequence, releases a transcript complementary to the CIS mRNA sequence. The TRANS/CIS <b>RNA duplex</b> prevents ribosomes from binding to RBS on target mRNA, thus <b>repressing protein synthesis</b>. The amount of the TRANS-repressor regulates the rate of translation of the target mRNA (<i>see Fig. 1, left panel</i>) | ||
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- | [[Image:project3b.png|center|950px|thumb|<center>Figure 1 - T- | + | [[Image:project3b.png|center|950px|thumb|<center>Figure 1 - T-REX device</center>]] |
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= Acknowledgements = | = Acknowledgements = | ||
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Revision as of 20:43, 21 October 2009
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Project Summary
Our idea
The aim of our project is the design of a new device to control the synthesis of any protein of interest. This "general-purpose" standard device, implemented in E. coli, acts at the translational level to allow a switch in protein expression faster than transcriptional promoter regulation. We named this device T-REX (Trans Repressor of EXpression).
How T-REX works
The device consists of two new BioBricks:
CIS-repressing and TRANS-repressor sequences were designed by BASER software.
Transcription of the target gene yields a mRNA strand - containing the CIS-repressing sequence at its 5' end - available for translation into protein by ribosomes (see Fig. 1, right panel). Induction of the promoter controlling the TRANS coding sequence, releases a transcript complementary to the CIS mRNA sequence. The TRANS/CIS RNA duplex prevents ribosomes from binding to RBS on target mRNA, thus repressing protein synthesis. The amount of the TRANS-repressor regulates the rate of translation of the target mRNA (see Fig. 1, left panel)
How we can test the device
In order to test and characterize our T-REX device, we developed the following genetic circuit (Fig 2):
More details about our work are reported in the Project section.
Acknowledgements
- [http://www.unibo.it/Portale/default.htm University of Bologna]
- [http://serinar.criad.unibo.it Ser.In.Ar. Cesena]
- Cultural Association San Sebastiano