BACTERIAL GENETICS

BACTERIAL GENETICS
Prokaryotes
Prokaryotes
the genome of prokaryotes is not in a
separate compartment: it is located in the
cytoplasm
prokaryotes contain no membrane-bound
organelles; their only membrane is the
membrane that separates the cell from the
outside world
nearly all prokaryotes are unicellular
Nucleoid
Nucleoid
the nucleoid is an irregularly-shaped
region within the cell of prokaryotes
where the genetic material is localized
the genome of prokaryotic organisms
generally is a circular, double-stranded
piece of DNA, of which multiple copies
may exist
Nucleic
Nucleic
acids
acids
structure
structure
backbone polynucleotide strands
consists of sugar+base+phosphate
sugar: ribose-RNA
2-deoxyribose-DNA
5 heterocyclic bases: holds N atom
adenine, guanine-purines
uracile, cytosine, thymine-pyrimidines
Hydrogen bonds between bases
2 A-T
3 C-G
Primary-single strand
Secondary-double helix
Tertiary-folding of double helix
DNA Replication
synthesis of new strands of DNA (precise
complementary copy) are established by
DNA polymerases (Taq polymerase)
in bacteria starts at fixed points-oriC
elongation of strand by addition of
nucleotides to 3OH end (5 to 3 end direction)
starts with primers (oligonucleotides)
replication fork
replication replication
Bacterial genetic information is stored
in
Chromosome
Plasmid
Bacterial
Bacterial
chromosome
chromosome
usually double stranded circular DNA
(Borrelia-linear chromosome)
condensed into nucleoid (supercoiled loop
domains, histon-like proteins)
no nuclear membrane
size from 0,56 x 10
6
bp Mycoplasma
genitalium
4,639 x 10
6
bp Escherichia coli
replication starts from specific site-origin of
replication (OriC)
Bacteria Chromosome(s)
Bacillus subtilis One circular 4,2 Mb
Brucella melitensis Two circular (2,1+1,2
Mb)
E. coli One circular 4,6 Mb
Vibrio cholerae Two circular (2,9+1,1
Mb)
Bacterial
Bacterial

chromosome
chromosome
Bacterial genes
short (average 1000 nucleotides)
biosynthetic genes
repetetive sequences
catabolic genes
loss of gene activity leads to loss of the
ability to catabolise (use) the compound
drug resistance and bacteriophage genes
functionally linked genes are grouped into
operons
all the genes in a operon are transcribed
onto one large mRNA
Replication of bacterial
chromosome
replication starts from specific site-
origin of replication (OriC) and
proceeds bidirectionally into terminus
region terC
50 kbp per min (1 miscorporation for
every 10
10
nucleotides)
Plasmids
important features of bacterial genome
no essential for life of bacteria
conjugative (mobilizable) x
nonconjugative (horizontal transfer)
spread by partition between cells or by
conjugation between donor cell and
recipient cell
circular DNA 1.000-500.000 bp
Phenotypic properties carried by
plasmids
ANTIBIOTIC RESISTANCE-usually
enzymatic inactivation of the antibiotics
VIRULENCE FACTORS
Toxins
Adherence factors
PRODUCTION OF ANTI-MICROBIAL
AGENTS-antibiotic, bacteriocins
METABOLIC PATHWAYS
-
-

Pseudomonas sp. (catabolic activity for
toluene, salicylic acid)
Plasmid cloning-introduction of new
genes into bacteria
Plasmids are selftransferable by
conjugation (encode the sex-pilus
proteins and other transfer genes)
the transfer of genetic material between
bacteria through direct cell-to-cell
contact
conjugation is the main transfer of
antibiotic resistance
Conjugation
Conjugation
Transposons
jumping genes
move from one site to another
important in moving of antibiotic
resistance genes between chromosome
and plasmids
simple transposons only encode
information for transposition
compound transposons carry other
information (antibiotic resistance)
cause genomic instability resulting in
gene inactivation
Retrotransposons: copy and paste
(RNA intermediate)
DNA transposons: cut and paste (no
RNA intermediate)
Pathogenicity islands
large tracts of chromosomal DNA (up to 4 x
10
6
bp) which have been obtained from
another bacterial species by horizontal
transfer
usually contain the genes required for
virulence (toxins) and antibiotic resistance
typical examples are adherence factors,
toxins, iron uptake systems, invasion factors
and secretion systems
usually absent from those non-
pathogenic organisms of the same or
closely related species
1. PAIs carry genes encoding one or more
virulence factors: adhesins, toxins, invasins,
etc.
2. They are located on bacterial chromosome
or may be a part of a plasmid
3. They are flanked by direct repeats: The
sequence of bases at two ends are the same
5. PAIs carry functional genes, e.g. integrase,
transposase, or part of insertion sequences
Examples of microbes with PAI
E.coli:
EHEC-enterohemorrhagic E. coli (diarrhoea
and hemolytic-uremic syndrome after
colonization of the large intestine)
EPEC-enteropathogenic E. coli (watery
diarrhea after colonization of the small
intestine)
UPEC-uropathogenic E. coli
Integrons
expression cassettes for antibiotic
resistance genes (the type and number
may vary)
able to capture genes
have not repeated sequences
have integrase gene
Mutations
caused that bacteria are very variable
and highly adaptable
mutation=change within DNA sequence
bacteria mutate 1000x more frequently
than human cells
Types of mutation
point mutation (affect one base pair)
insertion and deletion of larger DNA
fragment (usually due to transposons)
can alter the function of a gene
amino acid substitution in protein
premature termination of protein
synthesis (nonsence)
gene over-expression (changed
regulation)
MRSA-mutation in PBP (penicillin
binding proteins)
ESBL-point mutation in beta-lactamase
Transfer of genetic information
Transformation
Conjugation
Transduction
Transformation
uptake of naked DNA
occurs in most bacteria and play role in
changing the phenotype (variation in
Neisseria gonorrhoeae)
Conjugation
encoded by plasmids
major mechanism of antibiotic
resistance transfer
Transduction
bacteriophage mediated
(bacteriophages are bacterial viruses)
transduction results in phage
conversion