Part:BBa_K371019

pdu(Propanediol utilization)BMC shell gene under Plac control

This composite consists of eight pdu bacterial microcompartment(BMC) structure gene following a natural lac promotor(BBa_R0010). These genes come from four basic parts,pduAB,pduJK,pduN and pduTUV, which consist of the natural RBS and gene separately. This part alone can assemble an empty BMC shell complex, which can be used as a platform to de novo make artificial organelles with different functions can be constructed. With enzymes or binding proteins inside the shell, different nanoreactors or nanoreservoirs can be produced.

Sequence and Features

Assembly Compatibility:

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

  Illegal BamHI site found at 1894
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 3236
  
• 1000
  COMPATIBLE WITH RFC[1000]

In the expression of protein, we use and unite the method mentioned in the previous papers. [see protocol] The bacterial was induced by 1 mM IPTG at 30 ℃ for 4 hours. The results was as follows.[the one without pduN was a negative control used to prove the function of pduN: in the growth process, the one without pduN tends to grow with more deposit implying its possible role in the vertex]

We use the ultracentrifuge to purify the proteins, with 48000 g speed, we get our targeted proteins in the pellet.

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