Difference between revisions of "Part:BBa K1111002"

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As a proof of principle we inserted the hya Promoter in front of BioBrick BBa_I746916 which encodes superfolded GFP. Then the induction was tested by transforming cells with the construct, innoculating them in media with different pHs and finally measure their GFP expression with a plate reader and looking at their fluorescence under the microscope.
 
As a proof of principle we inserted the hya Promoter in front of BioBrick BBa_I746916 which encodes superfolded GFP. Then the induction was tested by transforming cells with the construct, innoculating them in media with different pHs and finally measure their GFP expression with a plate reader and looking at their fluorescence under the microscope.
  
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[[Image: Hya-Promoter+GFP Map.jpg|thumb|500px|center|Figure 1: Our final construct, containing the hya promoter and the gene for superfolded GFP.]]
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[[Image:Team-EPFL-Lausanne Hya-Promoter+GFP Map.jpg|thumb|500px|center|Figure 1: Our final construct, containing the hya promoter and the gene for superfolded GFP.]]
  
 
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Revision as of 17:33, 2 October 2013

Hya promoter and Superfolded GFP

Introduction

The hya promoter is encoded in the genome of K-12 MG1655 E.Coli bacteria. Upon external acidification it is activated and induced transcribtion of hydrogenase isoenzyme 1. This enzyme can then catalyze reversible oxidation of hydrogen.

Figure 1: From http://biocyc.org/ECOLI/NEW-IMAGE?type=GENE&object=EG10468. How the hya promoter is induced

As a proof of principle we inserted the hya Promoter in front of BioBrick BBa_I746916 which encodes superfolded GFP. Then the induction was tested by transforming cells with the construct, innoculating them in media with different pHs and finally measure their GFP expression with a plate reader and looking at their fluorescence under the microscope.

Figure 1: Our final construct, containing the hya promoter and the gene for superfolded GFP.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 184
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 184
    Illegal NheI site found at 499
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 184
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 184
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 184
    Illegal AgeI site found at 26
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 543