Part:BBa_K2572032

TlpA39-pTlpA-VioABDE In iGEM, bacteria for human are popular, almost a thousand pages in all the wiki pages mentioned therapeutic bacteria therapeutic bacteria such as diagnostic bacteria and drug delivery bacteria. However, project for drug delivery bacteria, drugs were released by the lysis of bacteria, so bacteria are destroyed, but naked recombinant DNA will enter the human body, which may be taken up by the human microbiome. This is a thermosensitive part that can produce an intermediate of the antibiotic violacein(Shinoda K et al., 2007). We characterized it as effective in the capacity monitor. 2018.igem.org/Team:RDFZ-China/Measurement

As we compared the fluorescence of E. coli with an empty vector and E. coli with VioABDE inserted and expressed. Both strains were incubated for 12 hours with fluorescence and OD constantly monitored. We can see there is a major difference in f.u.,which means the high resource consumed by VioABDE.

Using VioABDE designed by Cambridge 2009, utilized from SHSBNU 2017, we found it was a good candidate for the drug release since the product should be a small molecule which can diffuse out without cell lysis. So, we connect VioABDE to the pTlpA promoter, followed by TlpA39. In a real situation, we can use ultrasound tissue heating, to de-repress the pTlpA promoter, and activate the production of violacein, which is a precursor of antiphlogosis.

Fig shows the Protovilacinic acid（PVA) production increase as temperature increase. Thermal Regulated expression of VioABDE Sequence and Features

Assembly Compatibility:

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  COMPATIBLE WITH RFC[10]
• 12
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  Illegal NheI site found at 1087
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  COMPATIBLE WITH RFC[23]
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  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 3872
  Illegal AgeI site found at 4068
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• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 5297
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Reference

Shinoda, K, et al. "Biosynthesis of violacein: a genuine intermediate, protoviolaceinic acid, produced by VioABDE, and insight into VioC function. " Chemical Communications 41.40(2007):4140-4142.