Difference between revisions of "Part:BBa K598016"
 
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<partinfo>BBa_K598016 short</partinfo>
 
<partinfo>BBa_K598016 short</partinfo>
  
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This is a basic part that contains a T7 promoter, a ribozyme part numbered 2.5, a GFP part and a T7 terminator.
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This is a basic part that consists of T7 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 ZYY2.png|center|thumb|600px| '''Figure 1''' Construction of T7-promoter+TPP Down-regulated Hammerhead Ribozyme 2.5 with Native RBS+GFP+T7-terminator. This part consists of 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 IPTG 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:T7-2.5.png|center|thumb|600px| '''Figure 1'''Working curve of BBa_K598016. The inhibition ratio is fluorescence intensity under given TPP concentrations compared to that of without TPP. Constructed plasmids were transformed into E. coli DH5acells 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 IPTG.
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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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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 21:25, 5 October 2011

T7-promoter+TPP Down-regulated Hammerhead Ribozyme 2.5 with Native RBS+GFP+T7-terminator

This is a basic part that consists of T7 promoter, thiamine pyrophosphate (TPP) ligand responsive hammerhead ribozyme numbered 2.5 with native RBS (AAGGAGAT), BBa_E0040 and BBa_B0015. (fig.1)

Figure 1 Construction of T7-promoter+TPP Down-regulated Hammerhead Ribozyme 2.5 with Native RBS+GFP+T7-terminator. This part consists of 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.


This device can respond to different TPP concentration and inhibit gene expression when induced by 1mM IPTG sensitively. The working curve is shown in fig.2. The inhibition ratio is the value of fluorescence intensity compared to that of without TPP.

Figure 1Working curve of BBa_K598016. The inhibition ratio is fluorescence intensity under given TPP concentrations compared to that of without TPP. Constructed plasmids were transformed into E. coli DH5acells 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 IPTG.


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:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 171
    Illegal BamHI site found at 984
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


For the full characterisation of the device, please refer to BBa_K598003