Team:WITS-CSIR SA/Project/Parts
From 2011.igem.org
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Parts Submitted
Click on the part number to go to the associated registry page. Click on the part name for a brief description of the part.
Part Number | Part Name | Part Length |
---|---|---|
BBa_K537001 | Theophylline riboswitch (type 1) + CheZ | 698bp |
BBa_K537002 | Theophylline riboswitch 2 + CheZ | 702bp |
BBa_K537000 | Atrazine riboswitch | 730bp |
BBa_K537003 | Theophylline riboswitch (type 1) Venus C-fusion | 780bp |
BBa_K537004 | Theophylline riboswitch (type 2) Venus C-fusion | 781bp |
BBa_K537005 | Reporter mCherry (mRFP1) C-Fusion | 679bp |
BBa_K537006 | Venus C-fusion | 721bp |
BBa_K537007 | RBS CheZ N-fusion | 658bp |
BBa_K537008 | Atrazine Riboswitch-mRFP | 658bp |
BBa_K537009 | Promoter-Theophylline riboswitch 1-Venus-Double terminator | 960bp |
BBa_K537010 | Promoter-Theophylline riboswitch 2-Venus-Double terminator | 961bp |
BBa_K537011 | Promoter-Theophylline riboswitch 1-CheZ-Venus-Double terminator | 1583bp |
BBa_K537012 | Promoter-Theophylline riboswitch 2-CheZ-Venus-Double terminator | 1587bp |
BBa_K537013 | Promoter-RBS-CheZ-Venus-Double terminator | 1543bp |
BBa_K537014 | Promoter-Atrazine riboswitch-mCherry-Double terminator | 1543bp |
BBa_K537015 | Promoter-Atrazine riboswitch-CheZ-mCherry-Double terminator | 1543bp | BBa_K537016 | Promoter-RBS-CheZ-mCherry-Double terminator | 1543bp |
This DNA part will encode for an RNA riboswitch senstive to theophylline. The RBS is not exposed in the absence of theophylline. When theophylline is present, it binds to the riboswitch and causes a conformational change which leads to the exposure of the RBS and consequently translation of the adjoining gene. In this case, the gene which will be expressed is CheZ, which is a protein fundamental to bacterial movement. This theophylline riboswitch (type1) CheZ fusion BioBrick activates the expression of the CheZ gene in a theophylline-dependent fashion. It is composed of a theophylline-sensitive riboswitch clone 8.1 (Topp and Gallivan JACS 2007; BBa_K249026 and BBa_K411001) that is detached from its associated translation unit (coding region) and fused to a CheZ gene which has a N-fusion prefix and lacks a stop codon (although a TAG stop codon is provided by the RFC 25 suffix). The riboswitch regulated N-terminal part can be fused to a reporter or other CDS.
CheZ is the chief regulator of the molecular events that lead the counter clockwise rotation of the flagella motor during the chemotaxis signal transduction pathway of E.coli. This counter clockwise flagella motor rotation results in bacterial swimming (instead of tumbling) in the presence of a chemoattractant (in this case, theophylline).
This DNA part will encode for an RNA riboswitch senstive to theophylline. When no theophylline is present, the RBS within the riboswitch sequence is not exposed to translation machinery. When theophylline is present, it binds to the riboswitch and causes a conformational change which results in the RBS being exposed. This allows for the translation of the adjoining gene. In this case, the gene which will be expressed is CheZ - a fundamental protein in the signalling cascade of bacterial chemotaxis.
This theophylline riboswitch (type2) CheZ fusion BioBrick activates the expression of the CheZ gene in a theophylline-dependent fashion. It consists of a newly improved theophylline riboswitch clone 12.1 (Lynch and Gallivan NAR 2009) that is detached from its associated translation unit (coding region) and fused to a CheZ gene which has a N-fusion prefix and lacks a stop codon (although a TAG stop codon is provided by the RFC 25 suffix). The riboswitch regulated N-terminal part can be fused to a reporter or other CDS.
CheZ is the chief regulator of the molecular events that lead the counter clockwise rotation of the flagella motor during the chemotaxis signal transduction pathway of E.coli. This counter clockwise flagella motor rotation results in bacterial swimming (instead of tumbling) in the presence of a chemoattractant (in this case, theophylline).