Difference between revisions of "Part:BBa K1031100"
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CapR is a transcriptional activator which belongs to NtrC family as DmpR. Sequence analysis of both capR gene and CapR protein reveals the significant similarity with DmpR, for their protein sequences are distinct at 9 residues only. | CapR is a transcriptional activator which belongs to NtrC family as DmpR. Sequence analysis of both capR gene and CapR protein reveals the significant similarity with DmpR, for their protein sequences are distinct at 9 residues only. | ||
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'''Structure''' | '''Structure''' | ||
| − | CapR has four domains. A domain is the signal reception domain, which undergoes conformational change when exposed to proper inducers. B domain is a linker, regulating the relative spatial position of A domain and C domain. C domain is the transcriptional activation domain. D domain contains a helix-turn-helix motif, which is capable of binding DNA sequence on Po promoter. | + | CapR has four domains. A domain is the signal reception domain, which undergoes conformational change when exposed to proper inducers. B domain is a linker, regulating the relative spatial position of A domain and C domain. C domain is the transcriptional activation domain. D domain contains a helix-turn-helix motif, which is capable of binding DNA sequence on ''Po'' promoter.('''Fig 1''') |
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| − | <img src="https://static.igem.org/mediawiki/igem.org/3/3b/Peking2013_DmpR_Figure4.png" | + | <img src="https://static.igem.org/mediawiki/igem.org/3/3b/Peking2013_DmpR_Figure4.png" style="width:600px; margin-left:170px" /> |
| + | <p style="text-align:center"><b>Fig 1</b> Structure of CapR protein. From N terminal to C terminal are domain A, domain B, domain C, domain D. | ||
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'''Detection profile''' | '''Detection profile''' | ||
| − | CapR detects phenol and its’ derivatives such as catechol. The detection profile of CapR is widely overlapped with that of DmpR. Previous works showed that when exposed to inducers at the concentration of | + | CapR detects phenol and its’ derivatives such as catechol. The detection profile of CapR is widely overlapped with that of DmpR. Previous works showed that when exposed to inducers at the concentration of 100µM, CapR's response to phenol and catechol could reach 300 and 100 fold respectively via firefly luciferin assay. |
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<partinfo>BBa_K1031100 SequenceAndFeatures</partinfo> | <partinfo>BBa_K1031100 SequenceAndFeatures</partinfo> | ||
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| + | == '''Construction of biosensor circuit''' == | ||
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| + | ''Po''/CapR biosensor circuit is constructed. The coding sequence of CapR was synthesized. A ''Pc'' library of constitutive promoters are constructed to fine-tune the biosensor, which consists of ''Pc'' promoters <html><a href="https://parts.igem.org/Part:BBa_J23113">BBa_J23113</a></html> and <html><a href="https://parts.igem.org/Part:BBa_J23109">BBa_J23109</a></html> ('''Fig 2'''). | ||
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| + | <img src="https://static.igem.org/mediawiki/igem.org/thumb/a/a3/Peking2013_part_CapR-tt.png/800px-Peking2013_part_CapR-tt.png" style="width:600px; margin-left:170px" /> | ||
| + | <p style="text-align:center"><b>Fig 2</b> Schematic diagram of biosensor circuit Po/CapR. The orange arrowheads represent promoters. Green ovals stand for RBS(Ribosome Binding Site). Terminators B0015 are shown in red squares. | ||
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Latest revision as of 21:28, 27 September 2013
CapR-Terminator
Characterization
Instruction
CapR is a transcriptional activator which belongs to NtrC family as DmpR. Sequence analysis of both capR gene and CapR protein reveals the significant similarity with DmpR, for their protein sequences are distinct at 9 residues only.
Structure
CapR has four domains. A domain is the signal reception domain, which undergoes conformational change when exposed to proper inducers. B domain is a linker, regulating the relative spatial position of A domain and C domain. C domain is the transcriptional activation domain. D domain contains a helix-turn-helix motif, which is capable of binding DNA sequence on Po promoter.(Fig 1)
Fig 1 Structure of CapR protein. From N terminal to C terminal are domain A, domain B, domain C, domain D.
Detection profile
CapR detects phenol and its’ derivatives such as catechol. The detection profile of CapR is widely overlapped with that of DmpR. Previous works showed that when exposed to inducers at the concentration of 100µM, CapR's response to phenol and catechol could reach 300 and 100 fold respectively via firefly luciferin assay.
Sequence and Features
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 86
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 283
Illegal NgoMIV site found at 564 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 790
Illegal SapI.rc site found at 1447
Construction of biosensor circuit
Po/CapR biosensor circuit is constructed. The coding sequence of CapR was synthesized. A Pc library of constitutive promoters are constructed to fine-tune the biosensor, which consists of Pc promoters BBa_J23113 and BBa_J23109 (Fig 2).
Fig 2 Schematic diagram of biosensor circuit Po/CapR. The orange arrowheads represent promoters. Green ovals stand for RBS(Ribosome Binding Site). Terminators B0015 are shown in red squares.