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Cyanide Biosensor

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Cyanide Biosensor. Southern Utah University. Kurt Whittemore, Ray Uchida, Greg ... Pterin-Dependent Oxygenolytic Cleavage to Ammonia and Formate in Pseudomonas ... – PowerPoint PPT presentation

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Title: Cyanide Biosensor


1
Cyanide Biosensor
Southern Utah University Kurt Whittemore,
Ray Uchida, Greg Thompson, Stacy Fobert, Luke
Allen, Kelsey Allen, Autumn Wassmuth
2
Goal
3
Motivation
  • Cyanide is toxic
  • Traditional chemical tests are difficult

Titration Distillation
Coloration
4
Past Cyanide Biosensors
  • Measurement of inhibited respiration
  • Measurement of byproducts from
  • Pseudomonas fluorescens (degrades
  • cyanide)

(Nakamura and Karube. 2003)
(Nakamura and Karube. 2003)
5
General Regulated System
TR Promoter TR Gene Promoter
Response
6
Implementation
  • Modify genes from Pseudomonas fluorescens

Section of Pseudomponas fluorescens genome we
dealt with (5.6 kb).
(Fernandez et al. 2004)
7
BioBricks
8
Useful Combinations
9
Final Part for Our Project
TR Promoter TR Gene CN Promoter
GFP
10
Procedure
  • Transform cells with GFP
  • Isolate P.fluorescens genes with PCR
  • Cut genes with proper restriction enzymes
  • Ligation
  • Transform more cells

11
Results of PCR
12
GFP BioBrick
13
Restriction Digests
Diagram modified from iGEM wiki
14
Limitations of Biosensor
  • Cyanide could kill sensing bacteria
  • Sensing bacteria can only detect free cyanide

15
Future Work
  • Successfully build plasmid as planned
  • Determine if cyanide nitrilase gene is
  • regulated
  • Determine correct transcriptional regulator
  • Extend cyanide detecting system with
    complexed cyanide detecting capabilities

16
Conclusion
Copy/paste in biology is harder than on computer
17
Acknowledgments
  • Chief Advisor Charlotte Rosendahl Pedersen
  • Advisors Bruce Howard, Colette Calmelet
  • Funding provided by SUUSA, UGRASP
  • Southern Utah University

18
Citations
1. Nakamura, H., Karube, I. (2003). Current
research activity in biosensors. Anal Bioanal
Chem, 377, 446-468. 2. Fernandez, R.F., Dolghih,
E., Kunz, D.A. (2004). Enzymatic Assimilation of
Cyanide via Pterin-Dependent Oxygenolytic
Cleavage to Ammonia and Formate in Pseudomonas
fluorescens NCIMB 11764. Applied and
Environmental Microbiology, 70, 121-128.
19
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