Part:BBa_K2407111

Cup1:express a metallothionein to enrich copper ions in yeast

Cup1 is a kind of metallothionein which can combine with copper ions in yeast. we express this gene in our S.C-Cu to improve the ability to combine with coppers ions.

Characterization

In 2017 iGEM, Tianjin team finds that CUP1 could also be used for biological enrichment of copper in Saccharomyces cerevisiae. We connect TEF promoter, CUP1 and ura3 terminator by overlap. Homologous recombination helps us to insert the fragments to Synthetic chromosome Ⅴ. Thus we complete our gene circuit, which are used to adsorb copper ion later.Engineered yeast cells are cultured in YPD liquid media for 24 hours, after that the 430mg/L ions solution is added to the media. Atomic absorption spectroscopy is used to measure the concentration of coppers ions in the supernatant at equal intervals. The results are plotted as the adsorption curve in the figure.

Fig 1. the concentration curve of copper ions

Figure clearly shows the change of the concentration of copper ions in the supernatant. The concentration of cadmuim ions declines over time and S.C-Cu(genetically-engineered yeast) is more capable of binding copper ions than control group(BY4741). 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
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]