CaB Research
Leaders Challenge
Nathan Battersby, Gemma Mancuso, Murad
Jabarov, Viktoria Weisz and Ramya Selladurai
How can we improve Canada’s Natural
Resource sector?
Athabasca Oil
Sands, Alberta
Tailing Ponds
http://goo.gl/ORUzVo
Alcanivorax borkumensis
Active Genes:
ALK B1
ALK B2
Cytochrome P450
http://goo.gl/T7cMeS
Research
- Gene Knockout Techniques
-
Replacement of each gene
- Isolate the enzyme encoding genes to
test hydrocarbon-degrading capabilities
Canada meets high global
demand for oil by extracting
and refining Bitumen
http://goo.gl/7P75xB
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, 2009 43, 266-71.
[2] Canadian Association of Petroleum Producers. Oil Sands Today, 2015.
http://goo.gl/z96Cks
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
http://goo.gl/6WZoSu
[3] McNeill et al. Ecotoxicology and Environmental Safety, 2012 84, 254-261
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
Control!
Alkanivorax borkumensis has 3
genes Alk B1, Alk B2,
Cyt-P450 which will be tested
individually, all together
and in combinations
http://goo.gl/NpstbR
Homologous Recombination
=
The Gene Switch
The Products
Αlk Β1
Αlk Β2
Cyt - P450
Αlk Β1,
Αlk Β2 & Δ
Cyt-P450
the control
Biological Photocopier PCR
Polymerase Chain
Reaction
http://goo.gl/7f7HEe
http://goo.gl/n9pc8f
Amplification!
Replicate, replicate and replicate!
30 Cycles =
1,073,741,842 Copies!!
http://goo.gl/MS1rEX
Nucleic Acid Electrophoresis
http://goo.gl/8GAaie
http://goo.gl/YDbUov
DNA sequencing
http://goo.gl/vEhhjq
http://goo.gl/F8n4nd
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
➔ Future Research:
1) Developing Biofilm composed of A. borkumensis
2) 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
http://goo.gl/nIUofF
[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.
Acknowledgements
Alex Somerville
Rehman Ata
Daniel Prete
Crystal Gadishaw - Lue