Part:BBa_K5226074

B0064-porin281-Vib-ftl-porin194-Vib-folD

The existing methods for large-scale production of P34HB primarily rely on microbial fermentation. A key limiting factor in this process is the molar ratio of 4HB. Increasing the 4HB molar ratio can lead to a decrease in the melting temperature and apparent fusion heat of the copolymer, as well as an improvement in the polymer's deformation resistance. Therefore, enhancing the molar ratio of 4HB is crucial for the modification of P34HB.

Prior to embarking on this project, our laboratory had already conducted research on the production of P34HB. It was found that the expression of the 4hbd-sucD-ogdA-orfZ gene cluster could increase the molar ratio of 4HB. Following fermentation using Mmp1 inducible promoter, the porin194 constitutive promoter was considered more suitable based on the concentration gradient induction trend observed with IPTG.

Since two plasmids, pSEVA321 and pSEVA341, are commonly used in the laboratory, the gene cluster has only been previously expressed through the pSEVA321 plasmid. Our intention is to introduce the porin194-4hbd-sucD-ogdA-porin194-orfZ gene cluster into TD80 to synthesize P34HB, utilizing both the pSEVA341 and pSEVA321 plasmids, which allow us to evaluate which plasmid yields better results.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 1189
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 1189
  Illegal NheI site found at 2375
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 1189
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 1189
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 1189
  Illegal NgoMIV site found at 455
  
• 1000
  COMPATIBLE WITH RFC[1000]