Device

Part:BBa_K598011

Designed by: Yangyang ZHAO, Zhenrun ZHANG, Yan GONG   Group: iGEM11_Peking_R   (2011-09-27)
Revision as of 15:55, 5 October 2011 by Yana (Talk | contribs)

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. This device can respond to different TPP concentration and behave a sensitive response when induced by 1mM arabinose. And it mainly inhibit the gene expression. The working curve is shown in fig.1. The inhibition ratio is the value of fluorescence intensity compared to that of without TPP.

Figure 1Working 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 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 arabinose.

This part 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.

[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
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 125
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 65
    Illegal BamHI site found at 258
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 931


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Categories
Parameters
n/apBAD+TPP Down-regulated Hammerhead Ribozyme 2.5 with Native RBS+E0040+B0015