Part:BBa_K1197001

Anti MazF construct (pHyperspank + Anti MazF + Double Terminator)

Anti MazF is an antisense RNA producing sequence which is complemantary to MazF (K302033). MazF is a non-specific ribonuclease coding element which kills Bacillus subtilis. Anti MazF element produces an mRNA which is reverse complemantary to mRNA produced by MazF. Thus, Anti Mazf mRNA binds to MazF mRNA and inhibit its activity for coding MazF protein.

With pHyperspank promoter, the production of Anti MazF will be depend on IPTG presence in the medium. If there is IPTG in medium, it will inactivate LacI which inhibit pHyperspank promoter. Thus, Anti MazF will be produced. On the other hand, if there is no IPTG in medium, LacI will inhibit pHyperspank and Anti MazF mRNA cannot be produced. As a result MazF protein will be produced, and kills the bacteria.

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
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Gel electrophoresis photo of Anti MazF construct

Modelling Data

In the graph below, it is seen that when IPTG concentration is 0, LacI in the medium cause inhibition of pHyperspank. So we saw so little and decreasing amount of Anti MazF mRNA.

In the graph below, when IPTG=4000, LacI in the medium form complex with IPTG and the inhibition of it on pHyperspank decreases.