Transcript Ocean Viruses - The University of Arizona, Ecology and

From Isolates to Genomes
Tucson High School
Biotechnology Course
Spring 2010
Jennifer Brum
Originally from Las Vegas
BS in Marine Science (minor in Chemistry)
MS in Oceanography
PhD in Oceanography
Currently a post-doc in Dr. Matthew Sullivan’s lab at the University of Arizona
Mostly study aquatic viral ecology, but some biogeochemistry
Have conducted research in the North Pacific Ocean, Hawaii coastal regions,
Antarctica, Mono Lake (Sierras)
1 microliter (µl)
1 square mm (mm2)
1,000 bacteria
10,000 viruses
1 mouthful:
500 million viruses
50 million bacteria
Viruses are
host specific
Every living organism has viruses that infect it
… we just have to find them
Our Project:
Isolate and evaluate viruses that infect 3 marine cyanobacterial strains
Prochlorococcus sp.
Strain : MED4
Isolated from:
Mediterranean Sea
Adapted to high light
Prochlorococcus sp.
Strain : MIT9313
Isolated from:
Gulf Stream
Adapted to low light
Synechococcus sp.
Strain : WH7803
Isolated from:
Sargasso Sea
Virus Samples from Tara Oceans
Virus Samples from Tara Oceans
Italy
Spain
Mediterranean Sea
Africa
Virus Samples from Tara Oceans
The Process
1) Isolate viruses from Tara Oceans samples that infect
our 2 cyanobacteria cultures
2) Use PCR and DNA sequencing to
examine DNA sequences of specific
genes in the isolated viruses
- Hypotheses!
3) Look at isolated viruses with TEM
4) Analysis of sequenced viral genomes
from last year’s class
Today: Start the Isolation Process
Agarose Overlays
molten agarose (29 ºC)
cyanobacteria culture
virus sample
Plate
(side view)
Isolation
plaques
No Infection
Infection !!
The Plan
1/2 students –
Pro MED4 x 4 Tara Samples
1/2 students –
Syn WH7803 x 4 Tara Samples
33 students –
Each Tara sample gets screened 3 times
on each cyanobacteria strain
Replication!
Controls
cyanobacteria +
known virus
cyanobacteria +
no viruses
Positive Control
Negative Control
A bit about sterile technique
There are bacteria in the air
There are bacteria on your hands
There are bacteria in your hair
… There are bacteria everywhere!