work presentation oct 2015 .pdf
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Title: work presentation oct 2015
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INDUCIBLE TOXIN GENE
1. Staphylococcus aureus
2. S. aureus in cattle
3. Toxin - antitoxin systems
4. Lac Operon
6. Construction of gene cassette
7. Knockout cassette
8. Vaccine strategy
Gram positive coccal bacterium.
Immobile, forms grape like clusters as
they divide in two planes.
Facultative anaerobes, non-sporulating,
catalase positive, coagulase positive.
Colonies are yellow.
Reproduces asexually by binary fission.
Has about 33% G+C content and an
average gene length of 824 nucleotides
with 85% coding sequence.
Responsible for various diseases
including: mild skin infections, invasive
diseases, and toxin mediated diseases.
S. aureus also causes
economically important mastitis
in cows, sheep and goats.
Staphylococcus aureus IN CATTLE
Leading cause of contagious
mastitis in dairy cattle.
Severe damage to milk
producing tissue and leads to
Generally results in a herd
The bacteria persist in
mammary glands, teat canals,
and teat lesions of infected
cows. Mildly contagious.
Spread from cow to cow
primarily at milking time.
TOXIN-ANTI TOXIN SYSTEMS
LAC - OPERON
E. coli, S. aureus shuttle vector
CONSTRUCTION OF GENE CASSETTE
In S. aureus genome
MazF Up flank
MazF Dn flank
Extension by PCR
Insertion of fragment into
plasmid by Gateway technology
using BP clones