Part:BBa_K1111004

superfolded GFP driven by CAD promoter

This constructs expresses GFP when the pH is lowered and there is a high lysine concentration.

Introduction

The Cad Promoter was originally discovered in the K-12 E.coli substrain MG1655. It's original purpose is to induce Expression of Lysine decarboxylase. This Enzyme can remove carboxy Groups from acids, thus increasing the external pH of the medium. Thus the promoter is induced upon external acidification and excess Lysine and reaches Maximum Expression under anaerobic conditions.

Figure 1: From http://biocyc.org/ECOLI/NEW-IMAGE?type=GENE&object=EG10131. Mechanism that depicts the induction of the cad-promoter.

The idea then was to insert this promoter in front of an Enzyme that could degrade nanoparticle made out of gelatin in oder to release the nanonparticles Contents. As a proof of principle we inserted the CAD promoter in front of the bioBrick BBa_I746916 which encodes superfolded GFP.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 496
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 474
  Illegal SapI.rc site found at 540