Difference between revisions of "Part:BBa K598011"
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<partinfo>BBa_K598011 short</partinfo> | <partinfo>BBa_K598011 short</partinfo> | ||
− | This is a basic part that consists of pBAD promoter, thiamine pyrophosphate (TPP) ligand responsive hammerhead ribozyme, BBa_E0040 and BBa_B0015. | + | This is a basic part that consists of pBAD promoter, thiamine pyrophosphate (TPP) ligand responsive hammerhead ribozyme numbered 2.5 with native RBS (AAGGAGAT), BBa_E0040 and BBa_B0015.(fig.1) |
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+ | [[Image:PekingR ZYY1.png|center|thumb|500px| '''Figure 1''' Construction of pBAD+TPP Down-regulated Hammerhead Ribozyme 2.5 with Native RBS+E0040+B0015. This part consists of pBAD promoter, TPP ribozyme 2.5 with native RBS (AAGGAGAT), BBa_E0040 and BBa_B0015.It is obtained by PCR from inactive TPP-HHAz 2.5 [1] which is kindly provided by Markus Wieland ''et al.'', and then inserted into pSB1C3 through standard assembly. ]] | ||
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+ | This device can respond to different TPP concentration and inhibit gene expression when induced by 1mM arabinose sensitively. The working curve is shown in fig.2. The inhibition ratio is the value of fluorescence intensity compared to that of without TPP. | ||
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+ | [[Image:PEKING-R PBAD-2.5.png|center|thumb|500px| '''Figure 2'''Working curve of BBa_K598011. The inhibition ratio is fluorescence intensity under given TPP concentrations compared to that of without TPP. Constructed plasmids were transformed into E. coli DH5acells and characterized in M9 medium with a TPP concentration gradient of 0.001, 0.003, 0.01, 0.03, 0.1, 0.3, 1, 3 uM with being induced by 1mM arabinose. | ||
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+ | == Reference == | ||
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+ | [1] Markus Wieland, Armin Benz, Benedikt Klauser, and Jörg S. Hartig. (2009). Artificial Ribozyme Switches Containing Natural Riboswitch Aptamer Domains. Angew. Chem. 121, 2753-2756 | ||
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Latest revision as of 21:12, 5 October 2011
pBAD+TPP Down-regulated Hammerhead Ribozyme 2.5 with Native RBS+E0040+B0015
This is a basic part that consists of pBAD promoter, thiamine pyrophosphate (TPP) ligand responsive hammerhead ribozyme numbered 2.5 with native RBS (AAGGAGAT), BBa_E0040 and BBa_B0015.(fig.1)
This device can respond to different TPP concentration and inhibit gene expression when induced by 1mM arabinose sensitively. The working curve is shown in fig.2. The inhibition ratio is the value of fluorescence intensity compared to that of without TPP.
Reference
[1] Markus Wieland, Armin Benz, Benedikt Klauser, and Jörg S. Hartig. (2009). Artificial Ribozyme Switches Containing Natural Riboswitch Aptamer Domains. Angew. Chem. 121, 2753-2756
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 125
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 65
Illegal BamHI site found at 258 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 931