Difference between revisions of "Part:BBa K494004"
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In general we want to provide a new, scalable principle of gene regulation based on termination and antitermination which can be further developed, tested and optimizted by everybody. To enable that in the easiest way possible we decided to not just add the switches we designed but instead the material needed for evaluation. Therefore we focus on providing the parts needed for verification and testing of new individual switches. <br><br> | In general we want to provide a new, scalable principle of gene regulation based on termination and antitermination which can be further developed, tested and optimizted by everybody. To enable that in the easiest way possible we decided to not just add the switches we designed but instead the material needed for evaluation. Therefore we focus on providing the parts needed for verification and testing of new individual switches. <br><br> | ||
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| − | + | This is a ready to use part consisting of our screening backbone <partinfo>BBa_K494001</partinfo> and a His-terminator in front of mCherry for testing reasons. Following mCherry a short DNA sequence is encoded (the signal) which is transcriped into a short transmitter RNA which binds to a certain region of the His-terminator leading to antitermination. This signal is IPTG inducible, so in theory the terminator can be switched from "off" to "on" by an input.<br> | |
| − | + | This part belongs to the screening system for evaluation of individual switches constructed on the principle of bioLOGICS, in this case BBaK494004 may serve as a reference to compare results with those presented by the TU Munich 2010 iGEM Team. Together with <partinfo>BBaK494003</partinfo> (Terminator only) and <partinfo>BBaK494002</partinfo><partinfo> (positive control for mCherry expression) it is a setup to evaluate termination and antitermination efficiency of the His-Terminator. <br> | |
| − | This is a ready to use part consisting of our screening backbone BBa_K494001 and a His-terminator in front of mCherry for testing reasons. Following mCherry a short DNA sequence is encoded (the signal) which is transcriped into a short transmitter RNA which binds to a certain region of the His-terminator leading to antitermination. This signal is IPTG inducible, so in theory the terminator can be switched from "off" to "on" by an input.<br> | + | |
| − | This part belongs to the screening system for evaluation of individual switches constructed on the principle of bioLOGICS, in this case BBaK494004 may serve as a reference to compare results with those presented by the TU Munich 2010 iGEM Team. Together with BBaK494003 (Terminator only) and BBaK494002 (positive control for mCherry expression) it is a setup to evaluate termination and antitermination efficiency of the His-Terminator. <br> | + | |
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In theory upon induction with IPTG the signal will be transcripted into transmitter RNA which binds to the Trp-terminator, leading to antitermination and therefore allowing transcription of mCherry which can be detected as a rise of red fluorescence. Other PoPS devices can be tested in a similiar way based on the K494001 backbone and K494004 may serve as a reference then. | In theory upon induction with IPTG the signal will be transcripted into transmitter RNA which binds to the Trp-terminator, leading to antitermination and therefore allowing transcription of mCherry which can be detected as a rise of red fluorescence. Other PoPS devices can be tested in a similiar way based on the K494001 backbone and K494004 may serve as a reference then. | ||
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The His-Terminator (which does not exist as an individual BioBrick since there are enough thoroughly tested Terminators yet in the registry) is based on an attenuator stem loop in front of the ''his-operon'' of ''Salmonella enterica''. The corresponding signal was constructed as decribed in the [http://2010.igem.org/Team:TU_Munich/Project TU Munich iGEM 2010 wiki]. | The His-Terminator (which does not exist as an individual BioBrick since there are enough thoroughly tested Terminators yet in the registry) is based on an attenuator stem loop in front of the ''his-operon'' of ''Salmonella enterica''. The corresponding signal was constructed as decribed in the [http://2010.igem.org/Team:TU_Munich/Project TU Munich iGEM 2010 wiki]. | ||
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| + | [[Image:K494004.png|500px|thumb|center|Emission spectra of induced and uninduced BBa_K494004 ; A: eGFP fluorescence ex: 501 nm, B: mCherry fluorescence ex: 587 nm]]<br> | ||
Revision as of 03:00, 28 October 2010
Exemplary insert for BBa_K494001 (His-Term and His-Sig)
In general we want to provide a new, scalable principle of gene regulation based on termination and antitermination which can be further developed, tested and optimizted by everybody. To enable that in the easiest way possible we decided to not just add the switches we designed but instead the material needed for evaluation. Therefore we focus on providing the parts needed for verification and testing of new individual switches.
This is a ready to use part consisting of our screening backbone BBa_K494001 and a His-terminator in front of mCherry for testing reasons. Following mCherry a short DNA sequence is encoded (the signal) which is transcriped into a short transmitter RNA which binds to a certain region of the His-terminator leading to antitermination. This signal is IPTG inducible, so in theory the terminator can be switched from "off" to "on" by an input.
This part belongs to the screening system for evaluation of individual switches constructed on the principle of bioLOGICS, in this case BBaK494004 may serve as a reference to compare results with those presented by the TU Munich 2010 iGEM Team. Together with BBa_K494003 (Terminator only) and BBa_K494002No part name specified with partinfo tag.