Part:BBa_K5323990

P_tac-RBS-AFE_0053-TT

Uses a strong promoter Ptac (BBa_K180000), a factor strong RBS(BBa_J34801), AFE_0053(BBa_K5323000) and TT(BBa_B0015). This part is an easy BioBrick.

Usage and Biology

This component is used to overexpress DGC in the engineering bacterium Acidithiobacillus ferrooxidans and participate in screening the most efficient cyclic diguanylate dynamic synthesis strains. We cloned the highly efficient DGC AFE_0053 in A. ferrooxidans genome into the expression vector pYDT to construct the highly efficient expression vector of DGC. Furthermore, the carrier is conjured into A. ferrooxidans, the concentration of biofilm is measured, and the most efficient dynamic regulatory gene of c-di-GMP is screened.

Fig 1 Acidithiobacillus ferrooxidans growth curve

It can be seen from the figure that Acidithiobacillus ferrooxidans grows slowly, and we speculate that overexpressed c-di-GMP may have a toxic effect on the growth of A. ferrooxidans.

Fig 2 Acidithiobacillus ferrooxidans pore plate biofilm formation column

Since it is previously known that IPTG can promote the significant expression of biofilm, after obtaining the growth characteristics of A. ferrooxidans, we directly measure the biofilm content in different time periods when the inducer IPTG is added to the strain. At the highest value of biofilm formation 36 hours, AFE_0053 formed the thickest biofilm, showing a difference from the control.

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 NgoMIV site found at 2256
  Illegal AgeI site found at 2419
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 1680