1. ISOLATION OF A NOVEL
MYCOBACTERIOPHAGE
Lizbeth Pérez Castro
Javier Zavala Ayala
Mónica Del Moral
Félix Valles
2. 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.
3. 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
4. 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
5. 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.
8. 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.
12. 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
14. 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
16. CONCLUSION
• Two mycobacteriophages were found:
1. Nitidusvenutus
a. located in Carretera 943 Sector Los Chinos, Gurabo
b. ……. Were completed
2. Musamodel
a. located in 1-9 calle Barcelona, Urb. Terralinda, Caguas P.R
b. Up to now, we believe it has a lysogenic life cycle because of turbidness
c. three purifications were completed
17. 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