Part:BBa_K1011001

P + RBS + ftsZ + RBS + GFP + Term

This biobrick enables the complete production of ftsZ and GFP (as an indicator of production). The same RBS is used upstream of both ftsZ and GFP in order to approximate a 1:1 production, so that the flourescent intensity will give a relative indication of ftsZ production.

As this picture below illustrates, under normal WT conditions, FtsZ will form the Z ring at the middle of the cell. When overexpressed, FtsZ will polymerize non specifically at the polar ends of the cell, thereby cleaving the cell to form a minicell.

This biobrick is constructed with a GFP reporter gene downstream of the ftsZ under the control of the same promotor but with its own RBS. The purpose is to be able to have a visual confirmatin that the mRNA transcribed at least to produce GFP. The GFP would also make it easier to visualize the minicells underneath a flourescent microscope. As shown below, you can clearly see both minicells and rod shaped E. coli that are starting to cleave at the polar ends.

The graph below shows the cell count after 14 hours at different concentrations of the IPTG inducer. Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 403
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1903