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Phage
Plasmi
d
Transposo
1.PLASMID
Plasmid
Extrakromosom, benang ganda,
biasanya supercoiled
Ditemukan pada bakt G+ dan G- ;
satu sel individu sering harbors
membebaskan sejumlah plasmid
DNA adalah berada di luar e
kromosom; replikon plasmid
terpisah dari anakan sel dan
dipelihara di dalam sel anakannya
Plasmid manunjukkan sedikit
homologi (bila ada) thd sequen
kromosom DNA inang
Produksi bacteriocin
Degradasi seny2 toksik
Simbiosis dg jasad eukariot (fiksasi nitrogen)
Produksi antibiotik
Resisten thd antibiotik
Enterotoxins (E. coli)
Hemolysins
Mhslk Faktor adherence & colonization (pili)
Mhslk proteins serum resistensi
Mhslk patwai katabolik dan anabolik
Types of Plasmids
Fertility plasmids--F plasmids
(mating/conjugation).
Resistance plasmids--R plasmids
(antibiotic resistance--see handout)
Col plasmids--encode colicins
(bacteroicins)
Virulence plasmids--encode determinants
required for pathogenesis
Metabolic plasmids-mediate degradation
of toxic compounds
Insertionsequences(IS)assistinthe
integrationoftransposons(Tn)into
homologussitesofrecipientsgenome.
DifferentHfrstrainsareproducedasa
result
Genesforconjugativetransfer
Originoftransfer
Replication&segregationgenesofFplasmid
99kbF(Fertility)PlasmidGeneticMap(E.coli)
Understanding Plasmids
4. Types of plasmids & their biological significance
Remember- essential host functions not part of plasmid
genes
Pseudomonas- entire metabolic degradation pathways of
unique compounds camphor, napthalene.
Cryptic plasmids we know little of the plasmid functions
Understanding Plasmids
5. Resistance Plasmids (R plasmids)
R100 carries resistance for sulfonamdes, tetracyline,
chloramphenicol, spectinomycin, mercury. Broad enteric host
range- Escherichia, Proteus, Klebsiella, Salmonella, Shigella,
Understanding Plasmids
6. Toxins & other virulence characteristics
Ability to attach and colonise a host
o E. coli Colonisation Factor Antigen (CFA) assists in
attaching to intestinal epithelia
production of toxins, enzymes that damage the host
o E coli hemolysin- lyse RBS & E. coli enterotoxin- induce
salt and water secretion into the bowel
Understanding Plasmids
7. Bacteriocins
Bacterocins inhibit or kill related species or different strains of
the same species (limited inhibitory spectrum to antibiotics)
E. coli plasmids- Col
Plasmids: Examples
R plasmid
Panel A: coexistence of 2
plasmids from different
incompatibility (Inc) groups
-both maintained
Panel B: curing of one
plasmid. In the absence of
selection, asymmetric
partitioning results in the
loss of one of each plasmid
-ultimately cells
contain one
plasmids of the
other
but not both
ReplicasiPlasmid
Mengikuti aturan replikasi DNA kromosom, bila
replikasi mrpk stringent (low copy # plasmids;
e.g., F plasmids).
High copy # plasmids (i.e., relaxed plasmids)
Mengikuti aturannya sdr; mekanisme replikasi
distinct amplification of plasmid DNA
Bidirectional
Rolling circle
Plasmid transmission
Plasmids:Pathogenesis
A
Examples:Shigella(A)and
Yersinia(B)bothhavevirulence
plasmidsthatarerequiredfor
optimalvirulenceofthese
bacteria.Otherfactorsalso
contributetopathogenesisthat
arenotplasmidencoded(i.e.,
shigatoxin).Strainscuredof
theseplasmidshaveanattenuated
virulencephebnotype.
2. PHAGE
T4 dan T2 BAKTERIOFAG
Struktur Phage
1. Genome
single or double- benang DNA, sirkuler or linier.
linier, benang tunggal RNA.
2. Kapsid
protein shell ("coat").
pelindung asam nukleat genome.
Bentuknya icosohedral or filamentous
3. Ekor
Struktur protein yg nempel pd kapsid.
Struktur fisiknya sgt kompleks.
Terlibat dlm penempelan phage pd sel inang dan
pemindahan DNAphage ke sel inang
Tidak semua phages memiliki ekor
Generalised transduction
Lisogeni
Only observed for phage with double stranded
DNA genomes (phages capable of lysogeny are
called temperate).
Phage genome becomes part of host cell
chromosome (integrates into host DNA, e.g.
Lambda) or is maintained as a low copy number
plasmid (e.g. P1 phage).
Integrated phage genome called a prophage.
Bacterium with prophage called a lysogen.
Stages in Lysogeny
Adsorption
Introduction of phage DNA into host chromosome
(recombination-dependent event).
Expression: only of genes required for lysogeny.
Maintenance: prophage replicates along with bacterial
genome.
Induction: disruption of the lysogenic state and initiation of
the lytic phase.
Replication, Assembly and Release: as in the lytic cycle
Lytic Phages
RNA Phages (f2, MS2, R17, Q)
Simplest of phages.
Genome is single-stranded, linear RNA (3000-4000
bases), (+) strand.
Encode three proteins; CP (coat protein), A (attachment
protein), and Rep (RNA replicase).
All utilize F-pili as receptors for attachment.
Rep copies (+) strand into (-) strands then uses (-)
strands to make new (+) strands for packaging.
Lyse cells and release 10-20,000 phage per cell;
mechanism of lysis unknown.
Phages Litik
Single Stranded DNA Phages
T4
2. Genome Structure
T4
3. Life Cycle
-Infection with T4 causes shut-off of host
cell protein, DNA, and RNA synthesis.
-Progeny phage assembled within 15-20
min.
-Cells lyse within 20-25 min releasing
300 progeny phage.
Lambda ( )
Immunity:
Lysogenic strains of bacteria cannot be
infected with a phage of the same type as the
prophage.
Resistance to superinfection
In the lysogen, the cI repressor is dominant,
so an incoming phage is immediately
repressed and cannot replicate.
3.TRANSPOSON
Transposons
41
Transposable elements in
prokaryotes
Transposons (Tn)
Bacteriophage Mu
42
44
45
Conservative transposition
Replicative transposition
Retrotransposition
46
Conservative transposition
48
Replicative transposition
49
Retrotransposition
50
51
52
Transposons (Tn)
2 types of transposons:
Composite transposons
Noncomposite transposons
53
Composite
transposons
54
Insertionsequences(IS)assistinthe
integrationoftransposons(Tn)into
homologussitesofrecipientsgenome.
DifferentHfrstrainsareproducedasa
result
Genesforconjugativetransfer
Originoftransfer
Replication&segregationgenesofFplasmid
99kbF(Fertility)PlasmidGeneticMap(E.coli)
Composite Transposons