Part:BBa_K1111002

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