phage presentation spring 2014-2

15
ISOLATION OF A NOVEL MYCOBACTERIOPHAGE Lizbeth Pérez Castro Javier Zavala Ayala Mónica Del Moral Félix Vallés

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Page 1: Phage presentation  spring 2014-2

ISOLATION OF A NOVEL MYCOBACTERIOPHAGE

Lizbeth Pérez Castro

Javier Zavala Ayala

Mónica Del Moral

Félix Vallés

Page 2: Phage presentation  spring 2014-2

WHAT IS A BACTERIOPHAGE?

• A bacteriophage is any virus that infects any bacterium.

• Mycobacteriophages are viruses that infect mycobacteria.

• Found in different bacterial colonies.

• Mostly found in the soil and sea water.

• Resistant to antibiotics.

http://news.bbcimg.co.uk/media/images/66413000/jpg/_66413229_m0900092-bacteriophage_virus-

spl.jpg

Page 3: Phage presentation  spring 2014-2

BACTERIAS FOR HOSTING

1. M. smegmatis (Smeg)

• fast growing rate and non-pathogenic behaviour

• Aerobic organisms

• Found in water, soil and plants

2. Bacillus

• Obligate aerobes or facultative anaerobes

• Well understood- used as a model for differentiation, gene regulation, and cell cycle events in bacteria.

• Common in soil

Both are:

Gram positive

Acid fast bacteria

Morfology- rodshape

Page 4: Phage presentation  spring 2014-2

BACTERIOPHAGE STRUCTURE

• Phages consist of 3 basic components:

• Capsid-contains the genetic material

• Genetic material- DNA

• Tail-serves to attach to bacteria and DNA passes through tail and into bacterium

http://en.wikipedia.org/wiki/Bacteriophage

Page 5: Phage presentation  spring 2014-2

PHAGE LIFE CYCLES

• Temperate-Lysogenic:

• Replicate and lyse or dormant state- incorporate its genetic material into DNA of host.

• Virulent-Lytic:

• Straightforward cycle by lysing all bacteria they infect.

Page 6: Phage presentation  spring 2014-2

http://biomedfrontiers.org/infection-2014-3-28/

Page 7: Phage presentation  spring 2014-2

INTRODUCTION

• Problem: Can novel bacteriophages be isolated from tropical soil of Puerto Rico?

• Hypothesis: Unique bacteriophages will be isolated and characterized from a collected soil sample of Puerto Rico.

http://news.bbcimg.co.uk/media/images/66413000/jpg/_664

13229_m0900092-bacteriophage_virus-spl.jpg

Page 8: Phage presentation  spring 2014-2

PROCEDURE

httpwww.teachthechildrenwell.comscience.h

tml

http://www.exportersin

dia.com/swastikaelectr

ic/products.htm

http://quizlet.com/6760545/lab-3-streak-plate-

technique-flash-cards/

Soil

EnrichmentFiltration

Purification

HTPLTen Plate Preparation

Page 9: Phage presentation  spring 2014-2

MY BACTERIOPHAGE

• Name: Nitidusvenutus

• Located in: Carretera 943 Sector Los Chinos, Gurabo

• Coordinates: 18º 16’ 7’ N, 65º 58’ 32’ W

• Collected: February 24, 2014

• It was found in a rural area next to cow feces on a template day.

Page 10: Phage presentation  spring 2014-2

LOCATION

Page 11: Phage presentation  spring 2014-2

MY BACTERIOPHAGE

Page 12: Phage presentation  spring 2014-2

RESULTS

http://faculty.plattsburgh.edu/donald.slish/lambdatitre.html

Page 13: Phage presentation  spring 2014-2

CONCLUSION

• Two mycobacteriophages were found:

1. Nitidusvenutusa. Located in Carretera 943 Sector Los Chinos, Gurabo.

b. Based on the clear plaques, we can assume it has a lytic life cycle.

Page 14: Phage presentation  spring 2014-2

FUTURE PLANS

• Nitidusvenutus;

• Isolation of the bacteriophage’s DNA

• Bioinformatics analyses and sequencing.

Page 15: Phage presentation  spring 2014-2

ACKNOWLEDGEMENT

• Dr. Michael Rubin

• Juan Apiz

• Eduardo Correa

• Gustavo Martínez

• Giovanni Cruz

• Edwin Alvarado