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)

Every living organism has viruses that infect it … we just have to find them 1 microliter (µl) 1 square mm (mm 2 ) 1,000 bacteria 10,000 viruses 1 mouthful: 500 million viruses 50 million bacteria Viruses are host specific

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 X

Virus Samples from Tara Oceans

Spain Africa Italy Mediterranean Sea

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 No Infection Infection !! plaques

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 Positive ControlNegative Control cyanobacteria + known virus cyanobacteria + no viruses

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!