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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?
http://static.howstuffworks.com/gif/light-virus-1.jpg
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