Difference between revisions of "Part:BBa K4220100"
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==Why is this an improvement? == | ==Why is this an improvement? == | ||
| − | We have improved a kill switch in E. coli (composite part) into one which can function in the yeast by replacing each basic parts into ones which have similar functions in the yeast. | + | We have improved a kill switch in '''E. coli''' (composite part) into one which can function in the '''yeast''' by replacing each basic parts into ones which have similar functions in the yeast. |
Improvements: | Improvements: | ||
Latest revision as of 14:11, 1 October 2022
HSF1up-HIS3+NatR+ADH1+CUP1-HSF ORF
Introduction
This is a kill switch in yeast which can let the yeast survive only in the presence of copper ion.
This part is an improvement of part BBa_K2086002.
Why is this an improvement?
We have improved a kill switch in E. coli (composite part) into one which can function in the yeast by replacing each basic parts into ones which have similar functions in the yeast.
Improvements:
- promoter: PyeaR promoter (responsive to nitrate, nitrite and nitric oxide) to CUP1 (responsive to copper ion)
- essential gene: SerA (E. coli) to HSF1 (yeast)
Design
- How to kill the yeast?
- HSF1: essential gene in yeast [1]
- How to determine when to kill?
- Copper ion: abundant in the intestine; scarce in the environment [2]
- Others
- HIS3-NatR-ADH1: resistant gene expression for screening
Results
CUP1p-HSF1: kill switch
YPD: Yeast Extract Peptone Dextrose Medium
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 247
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 367
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 512
References
1 Kopczynski, J. B., Raff, A. C. & Bonner, J. J. Translational readthrough at nonsense mutations in the HSF1 gene of Saccharomyces cerevisiae. Mol Gen Genet 234, 369-378, doi:10.1007/BF00538696 (1992).
2 Nose, Y., Kim, B. E. & Thiele, D. J. Ctr1 drives intestinal copper absorption and is essential for growth, iron metabolism, and neonatal cardiac function. Cell metabolism 4, 235-244, doi:10.1016/j.cmet.2006.08.009 (2006).