Isolation and Characterization of Virus infecting Pyrodinium bahamenses from Mosquito Bay in Vieques

Luis A. Pérez SotoBiol 3009

University of Puerto Rico at CayeyRISE Program

Professor: Edwin Vazquez PhD

Dinoflagellates are unicellular organism that are part of the plankton an live suspended on the water surface and exhibits a great diversity of form.

The Pyrodinium bahamenses is the dinoflagellates that live on the Mosquitos Bay an is the one that is being infected by phages.

What are dinoflagellates?

This dinoflagellates are the responsible of the bioluminescent in the Mosquito Bay of Vieques.

http://bioartlab.blogspot.com/2008/11/se-han-conseguido-aisalar-proteinas.html

http://www.golden-heron.com/biobay.html

Are the most abundant organism in the biosphere and they are ubiquitous feature of the prokaryotic existence.

What are Virus?

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http://www.blogdosperrusi.com/wp-content/uploads/2010/02/virus.jpg

Virus

Isolate and characterize the virus that are infecting the Pyrodinium bahamenses to then know what are the effects of the infection.

Objetive

There is a lot of virus capable of infecting dinoflagellates.

Hypothesis

Direct extraction of the water with the organism.

It would be take 5 tubes in which everyone would have 15mL.

Identify the Dinoflagellate and the Phages

The identification of the microorganism would make by electron microscopy.

Methodology

Isolation of the Virus

To isolate the virus it will be created a cultive of P. bahamenses and then it would be added water with the virus. The virus will kill the P. bahamenses and what I am going to have on my substance is virus and dead P. bahamenses. After that the sample would be centrifugated and filtered to only have the virus on the sample.

Methodology

Sequence the DNAThe DNA would be sequenced by the Polymerase Chain Reaction (PCR).

After sequencing the DNA the data would be put on a database to see if the genome encoded with other genomes and with that we would be able to know who the virus is.

Methodology

Search on a Data Base if the genome is already characterize.

If its genome is not characterize I would use transposon mutagenesis of the Pyrodinium bahamenses host and identified the genes that are essential for the phages infection.

Methodology

By analysis of the mutation on the P. bahamenses I can determine some characteristic of the phage.

Methodology

The expected results would be that dinoflagellate is been infected by a virus which is causing a mutation on the dinoflagellate genome.

Expected Result

Eneida Diaz Ph.D. Edwin Vazquez Ph.D.

Acknowledgment

Clokie MR, Millard AD, Letarov AV, Heaphy S., Phages in nature, Pubmed.gov, January 2011 Recuperated from: http://www.ncbi.nlm.nih.gov/pubmed/21687533

Garbe J, Bunk B, Rohde M, Schobert M., Sequencing and Characterization of Pseudomonas auruginosa phage JG004., Pubmed.gov, May 14, 2011., Recuperated from: http://www.ncbi.nlm.nih.gov/pubmed/21569567

Julie E. Samson, Sylvain Moineau., Characterization of Lactococcus lactis Phage 949 and Comparison with other Lactococcal Phages., Pubmed.gov., August 27, 2010., Recuperated from: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2953028/?tool=pmcentrez

References

Ibarra Urruela Borja., Caracterizacion del Mecanismo de empaquetamiento in vitro del DNA del fago 029., mastesis.com., Year 2000., Recuperated from: http://www.mastesis.com/tesis/caracterizacion+del+mecanismo+de+empaquetamiento+in+vitro+de:87728

The Bioluminescent Bay or Biobay in Vieques http://www.golden-heron.com/biobay.html

References