Difference between revisions of "Part:BBa K1351015"
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<partinfo>BBa_K1351015 short</partinfo> | <partinfo>BBa_K1351015 short</partinfo> | ||
− | Quorum sensing two component system with native RBS and Terminator (histidine kinase comD, response regulator comE) of ''Streptococcus pneumoniae'' (R6) able to sense the competence stimulating peptide CSP. | + | Quorum sensing two component system with native RBS and Terminator (histidine kinase comD, response regulator comE) of ''Streptococcus pneumoniae'' (R6) able to sense the competence stimulating peptide CSP. CSP, a 17 amino acids long peptide pheromone, binds to and consequently activates the membrane-embedded sensor kinase ComD by changing the conformation of the polytopic kinase. ComD autophosphorylates and subsequently transphosphorylates the cognate response regulator ComE. ComE acts as transcription activator via binding to -10 promoters regions of early and late competence genes. These ComE binding sites (CEbs) consist of a 9 bp direct repeat separated by a stretch of 12 nucleotides [http://www.ncbi.nlm.nih.gov/pubmed/23216914 (Martin et al. 2013)]. [https://parts.igem.org/Part:BBa_K1351024 BBa_K1351024], [https://parts.igem.org/Part:BBa_K1351025 BBa_K1351025] are BioBricks containing CEbs form different promoters of ''S. pneumoniae''. |
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+ | [[File:LMU14 ComEdep promoters.png|600px]] | ||
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+ | ComE dependent promoters containing bases matching (blue) and differing (red) from the CEbs consensus sequence. (figure modified from [http://www.ncbi.nlm.nih.gov/pubmed/23216914 Martin et al. 2013)] | ||
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+ | In contrast to [https://parts.igem.org/Part:BBa_K1351016 BBa_K1351016] the native RBS was maintained, and no optimized RBS for ''Bacillus subtilis'' was added. | ||
<br><br>Used oligonucleotides | <br><br>Used oligonucleotides |
Revision as of 18:57, 17 October 2014
comDE Quorum Sensing two component system - native
Quorum sensing two component system with native RBS and Terminator (histidine kinase comD, response regulator comE) of Streptococcus pneumoniae (R6) able to sense the competence stimulating peptide CSP. CSP, a 17 amino acids long peptide pheromone, binds to and consequently activates the membrane-embedded sensor kinase ComD by changing the conformation of the polytopic kinase. ComD autophosphorylates and subsequently transphosphorylates the cognate response regulator ComE. ComE acts as transcription activator via binding to -10 promoters regions of early and late competence genes. These ComE binding sites (CEbs) consist of a 9 bp direct repeat separated by a stretch of 12 nucleotides [http://www.ncbi.nlm.nih.gov/pubmed/23216914 (Martin et al. 2013)]. BBa_K1351024, BBa_K1351025 are BioBricks containing CEbs form different promoters of S. pneumoniae.
ComE dependent promoters containing bases matching (blue) and differing (red) from the CEbs consensus sequence. (figure modified from [http://www.ncbi.nlm.nih.gov/pubmed/23216914 Martin et al. 2013)]
In contrast to BBa_K1351016 the native RBS was maintained, and no optimized RBS for Bacillus subtilis was added.
Used oligonucleotides
Prefix: GATCGAATTCGCGGCCGCTTCTAGAGGAAATAAGGGGAAAGAGTAATGG
Sufix: GATCTACTAGTAGCGGCCGCTGCAGATAGTCCGTACGGGATTCG
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1183
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]