Part:BBa_K3126019

nixA-KanR

NixA gene with GPM1 promoter and TDH1 terminator, using KanR as resistance cassette. We characterized the ability to adsorb nickel ions of Saccharomyces cerevisiae which expressed this part.

Usage and Biology

The sludge in the activated sludge process consists of a variety of microorganisms, which originally include yeast. Due to its excellent heavy metal tolerance, our project aim to develop engineering yeast to absorb nickel ions. The Composite part is used to homologous recombination in the chromosome of Saccharomyces cerevisiae.

In order to achieve this goal, we use NixA to transfer nickel ions into cells. NixA is a channel protein that can transfer nickel ions from the external environment into the cell’s internal environment [1]. Kanamycin is a resistance gene, and was used to screen successfully homologous recombination of genes. Under the coordination of these sets of genes, our engineered yeast can actively bind or absorb nickel ions, and its tolerance to nickel ions is greatly increased.

Result

Here is the curve comparing with original Saccharomyces cerevisiae. If you want to know more about our experimental methods, please click here https://2019.igem.org/Team:HBUT-China/Notebook .

Figure 1. Engineering yeast comparing with original yeast

Conclusion

The absorption abilities of engineered yeast and original yeast were compared, and the results indicated that among all of the engineering yeast, the S.cerevisiae/BBa_k3126019 (nixA-KanR) showed higher absorption efficiency than original yeast. Our results proved that this composite part is a biologically functional composite part.

Potential applications

In the future, this Composite part can be used to be introduced to other species of microorganisms to improve their nickel ion absorption capacity.

References

[1] Deng, X., He, J., & He, N. (2013). Comparative study on Ni2+-affinity transport of nickel/cobalt permeases (NiCoTs) and the potential of recombinant Escherichia coli for Ni2+ bioaccumulation. Bioresource technology, 130, 69-74.

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 2038
  
• 1000
  COMPATIBLE WITH RFC[1000]