Difference between revisions of "Part:BBa K4220100"

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This is a kill switch in yeast which can let the yeast survive only in the presence of copper ion.
 
This is a kill switch in yeast which can let the yeast survive only in the presence of copper ion.
  
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==How to kill the yeast? ==
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This part is an improvement of part [https://parts.igem.org/Part:BBa_K2086002 BBa_K2086002].
  
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HSF1: essential gene in yeast [1]
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==Why is this an improvement? ==
  
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==How to determine when to kill? ==
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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.
  
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Copper ion: abundant in the intestine; scarce in the environment [2]
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Improvements:
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*promoter: PyeaR promoter (responsive to nitrate, nitrite and nitric oxide) to CUP1 (responsive to copper ion)
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*essential gene: SerA (E. coli) to HSF1 (yeast)
  
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==Others==
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==Design==
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*How to kill the yeast?
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**HSF1: essential gene in yeast [1]
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*How to determine when to kill?
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**Copper ion: abundant in the intestine; scarce in the environment [2]
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*Others
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**HIS3-NatR-ADH1: resistant gene expression for screening
  
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HIS3-NatR-ADH1: resistant gene expression for screening
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==Results==
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[[File:neu100.jpg|550px|options|caption]]
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CUP1p-HSF1: kill switch
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YPD: Yeast Extract Peptone Dextrose Medium
  
 
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<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===
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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

caption

CUP1p-HSF1: kill switch

YPD: Yeast Extract Peptone Dextrose Medium

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 247
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 367
  • 1000
    INCOMPATIBLE 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).