Part:BBa_K3076803

Expression of CgMT driven by T7 promoter under LacO regulation

Description

This composite part was used to express the Corynebacterium glutamicum metallothionein gene (CgMT) in BL21(DE) E. coli strain. The expression of CgMT showed an increase in the copper absorption ability in E. coli which suits our purpose of creating a bacterial absorbent of metal pollutants in the liquid medium.

The CgMT coding sequence (BBa K3076100) was tagged with 6xHis tag and expressed under the regulation of the strong T7 promoter (BBa I719005), a lac operator (BBa K3076802) and a strong RBS (BBa K3076801). The terminator is T7 terminator (BBa K395601).

Usage and Biology

The E. coli expressing CgMT was put into copper (II) sulphate-containing medium. The concentration of copper ions in the mediunm were measured at different time points. We found that when compared with no IPTG induction control group and empty vector control group, the CgMT-expressing group removed significantly more copper ions inside the medium after 4 hours of incubation. This result indicates CgMT can effectively increase the copper absorption ability of E. coli and it serves the aim of our project which we tried to increase the metal pollutant removal ability of E. coli.

The CgMT<i>-expressing group removed ~30% of copper ions in the medium when compared with controls removing only ~12 - 15%.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal XbaI site found at 47
  Illegal PstI site found at 356
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 356
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal XbaI site found at 47
  Illegal PstI site found at 356
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal XbaI site found at 47
  Illegal PstI site found at 356
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 137