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| Construction of an Engineered Yeast with Green Fluorescent Protein Gene and Its Fluorescence in Response to Copper Ion |
| YI Min1,2, LÜ Pin1, WU Li-hua1,3, YI Hui-lan 1 |
1. School of Life Science, Shanxi University, Taiyuan 030006, China;
2. Department of Statistics, University of Missouri-Columbia, Columbia, MO 65211, USA;
3. Department of Biology, Taiyuan Normal University, Taiyuan 030031, China |
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Abstract The encoding region of green fluorescent protein (GFP) gene and the promoter region of S. cerevisiae metallothionein (CUP1) gene were amplified to construct a recombinant expression vector pCUP9K-GFP. By using lithium chloride transformation method, the recombinant expression vector was transformed into P. pastoris GS115 cells to create a yeast strain. Green fluorescent protein can be detected in engineered yeast cells and in their fermentation supernatant, indicating that engineered yeast cells could express GFP normally and then secrete GFP to the extracellular medium. The relative fluorescent intensity of the culture supernatant increased in a concentration-dependent manner after engineered yeast exposed to copper ions in a concentration ranging from 5μmol/L to 1mmol/L. However, the fluorescence of engineered yeast did not respond to chromium, cadmium and arsenic ions. This results suggest that CUP1 promoter can be induced specifically by copper ions, which may be used for the monitoring of environmental copper contamination.
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Received: 13 July 2012
Published: 25 October 2012
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