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ISOLATION OF A NOVEL MYCOBACTERIOPHAGE
Lizbeth Pérez Castro
Javier Zavala Ayala
Mónica Del Moral
Félix Valles
RISE Program UPR Cayey
WHAT IS A BACTERIOPHAGE?• A bacteriophage is any virus that infects any bacterium.
• Mycobacteriophages are viruses that infect mycobacteria.
• All mycobacteriophages are bacteriophages, but not all bacteriophages are mycobacteriophages.
• Found in different bacterial colonies.
• Mostly found in the soil and sea water.
• Resistant to antibiotics.
BACTERIAS FOR HOSTING1. 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
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
PHAGE LIFE CYCLES
• Temperate-Lisogenic:
• Replicate and lyse or dormant state- incorporate its genetic material into DNA of host.
• Virulent-Lytic:
• Straightforward cycle by lysing all bacteria they infect.
http://biomedfrontiers.org/infection-2014-3-28/
PROCEDURE
httpwww.teachthechildrenwell.comscience.
html
http://www.exportersindia.com/swastikaelectric/products.htm
http://quizlet.com/6760545/lab-3-streak-plate-technique-flash-cards/
Soil
EnrichmentFiltration
Purification
HTLTen Plate Preparation
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.
LOCATION
MY BACTERIOPHAGE
RESULTS
http://faculty.plattsburgh.edu/donald.slish/lambdatitre.html
MY BACTERIOPHAGE: MUSAMODEL
• Name- Musamodel
• Located in: 1-9 calle Barcelona, Urb.Terralinda, Caguas P.R
• Coordinates: 88˚14’6”N , 66˚2’39”W
• Collected: March 16, 2014
• Time: 7:51pm
• Temperature: 79˚
• About 3 inches deep and 1 foot close to concrete
• Found next to a plantain tree
• Soil was moist and loose
LOCATION
SAMPLES COLLECTED
1st 2nd 3rd 4th 5th 6th 7th
Date 2/6/14 2/18/14 2/24/14 3/3/14 3/10/14 3/16/14 3/16/14
Time 7:53 am 7:06 am 9:55 pm 9:38 am 6:26 am 7:45 pm 7:51 pm
Coordinates 18˚14’6”N, 66˚2’40”W
18˚14’13”N, 66˚2’35”W
18˚14’7”N, 66˚2’39”W
18˚14’18”N, 66˚2’42”W
18˚14’7”N, 66˚2’40”W
18˚14’6”N, 66˚2’39”W
18˚14’17”N, 66˚2’42”W
Temperature 75˚ 75˚ 79˚ 81˚ 73˚ 79˚ 79˚
Soil description
Moist, 3 inches deep
Moist, 1 inch deep
Moist, 1 inch deep
Moist, with moss, 1 inch deep
Moist, 1 inch
Moist, loose, 5 inches deep
Moist, loose, 3 inches deep
Site description
Urban, 5 feet close to concrete
Urban, 1 foot from concrete
Urban, 2 feet from concrete
Urban, ½ foot from concrete
Urban, 3 feet away from concrete
Urban, 1 foot close to concrete
Next to a plantain plant, 1 foot close to concrete
RESULTS
CONCLUSION
• Two mycobacteriophages were found:
1. Nitidusvenutusa. located in Carretera 943 Sector Los Chinos, Gurabob. ……. Were completed
2. Musamodela. located in 1-9 calle Barcelona, Urb. Terralinda, Caguas P.Rb. Up to now, we believe it has a lysogenic life cycle because of turbidnessc. three purifications were completed
FUTURE PLANS
• Nitidusvenutus
• Musamodel;• 3rd purification- to have more isolated plaques and measure diameter• Spot test• Make phage stocks-
• Empirical test• 10 plate preparation
• Analyze • Bioinformatics and sequencing
ACKNOWLEDGEMENT
• Dr. Michael Rubin
• Juan Apiz
• Eduardo Correa
• Gustavo Martínez
• Giovanni Cruz