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).
This composite part was synthesized by the ThermoFisher GeneArt platform. The CDS was codon-optimized for the expression in E. coli.
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.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 47
Illegal PstI site found at 356 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 356
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 47
Illegal PstI site found at 356 - 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 47
Illegal PstI site found at 356 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 137
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