Isolation and characterization of the phages that are infection the dinoflagellates from mosquito...
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Transcript of Isolation and characterization of the phages that are infection the dinoflagellates from mosquito...
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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
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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?
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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
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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
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http://www.blogdosperrusi.com/wp-content/uploads/2010/02/virus.jpg
Virus
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Isolate and characterize the virus that are infecting the Pyrodinium bahamenses to then know what are the effects of the infection.
Objetive
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There is a lot of virus capable of infecting dinoflagellates.
Hypothesis
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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
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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
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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
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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
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By analysis of the mutation on the P. bahamenses I can determine some characteristic of the phage.
Methodology
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The expected results would be that dinoflagellate is been infected by a virus which is causing a mutation on the dinoflagellate genome.
Expected Result
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Eneida Diaz Ph.D. Edwin Vazquez Ph.D.
Acknowledgment
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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
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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