1. CaB Research Leaders Challenge
Nathan Battersby, Gemma Mancuso, Murad Jabarov, Viktoria Weisz and Ramya Selladurai

2. How can we improve Canada’s Natural Resource sector?
Athabasca Oil
Sands, Alberta
Tailing Ponds

3. Alcanivorax borkumensis
Active Genes:
ALK B1
ALK B2
Cytochrome P450
[1] Teimoori, A., et al. (2012). Biotechnology and Applied Biochemistry 59(6):

4. Research Gene Knockout Techniques
Replacement of each gene
Isolate the enzyme encoding genes to test hydrocarbon-degrading capabilities

5. Canada meets high global demand for oil by extracting and refining Bitumen

6. Tailing Ponds
Water used in the hot water filtration process must be stored in tailing ponds due to its toxicity [1]
Contents of water in tailing ponds:
- sand
- clay
- residual bitumen [2]
Tailing ponds are located along the Athabasca River in Alberta
[1] Frank et al. Environmental Science & Technology, ,
[2] Canadian Association of Petroleum Producers. Oil Sands Today, 2015.

7. Rainbow Trout
Rainbow trout exposed to water influenced by tailings for short periods of time experience negative health effects [3]:
-Fin erosion
-Compromised immune systems
-Decrease in size of spleens
[3] McNeill et al. Ecotoxicology and Environmental Safety, ,

8. Hypothesis
The genes that encode for enzymes that degrade hydrocarbons found in our bacteria A. borkumensis and their success rate in degrading hydrocarbons found in tailing ponds

9. Control! Alkanivorax borkumensis has 3 genes Alk B1, Alk B2,
Cyt-P450 which will be tested
individually, all together
and in combinations

10. Homologous Recombination=
The Gene Switch

11. The Products Αlk Β1, Αlk Β2 & Δ Cyt-P450 Αlk Β1 Αlk Β2 Cyt - P450
the control

12. Biological Photocopier PCR
Polymerase Chain Reaction

13. Amplification! Replicate, replicate and replicate!
30 Cycles = 1,073,741,842 Copies!!

14. Nucleic Acid Electrophoresis

15. DNA sequencing

16. Conclusion
We aim to provide an ecologically friendly method to deal with the hydrocarbon waste products of the Athabasca Oil Sands operations
The bacterium A. borkumensis will be able to perform the task of hydrocarbon degradation
Laboratory techniques used:
Gene knockout, homologous recombination, gene knockout, polymerase chain reaction, nucleic acid electrophoresis

17. Developing Biofilm composed of A. borkumensis
Future Research:
Developing Biofilm composed of A. borkumensis
Determining exact byproducts of A. borkumensis metabolism
Bioengineering:
Replacement of Alk B1, Alk B2 and Cyc P450 with kanamycin resistance gene
Our Overall Goal:
Bioremediation of tailing ponds, in order to clean
up the environment
[9] Reva et al. Global features of the Alcanivorax borkumensis SK2 genome, 2007
[10] Sabriova et al. Functional genome analysis of Alcanivorax borkumensis strain SK2: Alkane metabolism , environmental adaptations and biotechnological potential.

18. Acknowledgements Alex Somerville Rehman Ata Daniel Prete
Crystal Gadishaw - Lue