Composite

Part:BBa_K1111002

Designed by: Luisa Spisak   Group: iGEM13_EPF_Lausanne   (2013-09-19)

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 2: 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


[edit]
Categories
Parameters
None