Microbial Genetics

Definitions
• Genetics
• the study of heredity, genes and the
mechanisms that they carry this
information
• Replication
• Expression
• Genome
• Complete genetic information of the cell
Definitions

• Chromosome
• The structures that are composed of DNA
that carry the hereditary information
• Gene
• Segments of the chromosome that code for
a specific product (usually a protein)
• Genomics
• Sequencing and molecular characterization
of genomes
Definitions
• DNA (deoxyribose nucleic acid)
• Nucleotides
• 3 components
• Phosphate
• Deoxyribose sugar
• Nitrogenous base
• Adenine, thiamine, cytosine or guanine
• Double helix (complementary strands)
• Base pairs
• A-T
• C-G
• A-U (RNA)
• Hydrogen bonds
DNA to proteins

• Gene on DNA

• Converted to mRNA

• mRNA on ribosome

• tRNA brings amino acids to ribosome for

protein synthesis
Definitions

• Genotype
• Genetic information of the organism
• Information that codes for characteristics of
the organism
• Phenotype
• The expressed or physical characteristics of
the organism
• The expression of the genotype
Bacterial Chromosome (DNA)

• Bacterial chromosome
• Single
• Circular
• Attached one or many
sites to plasma
membrane
Bacteria chromosome

• Escherichia coli
• 4.6 million base pairs
• 4300 genes
• 1mm long
• 1,000 X length of cell
• Supercoiled
• Topoisomerase II
• DNA gyrase
DNA replication

• Parental strand
• Two new “daughter
strands”
• Each strand acts as
template for new
strands
• Semiconservative
replication
RNA Synthesis

• Transcription
• Process of taking DNA code and
converting to RNA code
• Translation
• Converting RNA (mRNA) with tRNA to
form amino acid sequences and proteins
• Occurs at ribosome
Genetic Transfer and Recombination
• Genetic recombination
• Exchange of genes between
two DNA molecules to form
new combinations of genes on
a chromosome
• Crossing over
• Two chromosomes break and
rejoin
• Adds to genetic diversity
Genetic transfer and recombination
• Eukaryotes
• Meiosis
• Prophase I
• Prokaryotes
• Numerous different ways
Genetic Transfer and Recombination
• Vertical gene transfer
• Genetic information passed from an organism to its offspring
• Plants and animals
• Horizontal gene transfer
• Bacteria transfer genetic information form one organism to another in the
same generation
• Genetic information passed laterally
Horizontal Gene Transfer

• Horizontal gene transfer

• Donor cell
• Organism gives up its entire DNA
• Part goes to recipient cell
• Part is degraded by cellular enzymes
• Recipient cell
• Receives portion of donor cells DNA
• Incorporates donor DNA into its own DNA
• Recombinant DNA
Transformation

• Genes transferred from one bacterium to another

in solution
• Naked DNA
• Discovered by Griffith
• Used Streptococcus pneumoniae
• Two strains
• Virulent (pathologic) strain
• Had a polysaccharide capsule resists phagocytosis
• Avirulent (non- pathogenic) strain
• Lacked a capsule
Griffith’s Experiment
Transformation

• Bacteria after cell death and lysis could release DNA

into environment
• Recipient cell can take up DNA fragments and
incorporate into their own DNA
• Resulting in a hybrid (recombinant cell)
• Recombinant cell must be competent
• Able to alter cell wall to allow DNA (large molecule) to enter
• Bacillus, Haemophilus, Neisseria, Acinetobacter, and some Staph
and Strep
Genetic Transformation
Conjugation

• Conjugation
• Involves plasmid
• Circular piece of DNA
• Replicates independent of
chromosome
• Non essential for growth genes
• Requires cell to cell contact
• Opposite mating type
• Donor cell carries plasmid
• Recipient cell lacks plasmid
Conjugation

• Gram positive
• Sticky surfaces cause
bacteria to come in
contact with one another
• Gram negative
• Utilize sex pili
Conjugation

• E coli model
• F factor plasmid
• Fertility factor
• Donors (F+)
• Recipients (F-)
• Converted to (F+)
• F+ factor integrated into
chromosome
• Becomes Hfr (high
frequency of recombination)
cell
Bacterial Conjugation

• Hfr conjugates with F- cell

• Chromosomal strand
replicates and transferred to
recipient
• Incomplete transfer of donor
DNA
• Recipient integrates new
DNA
• Acquires new versions of
chromosome
• Remains F- cell
Conjugation in E. coli
Conjugation

• Minutes and conjugation

• Identify locations of
various genes
• Hfr
• His, pro, thr, leu, and F (+)
• F(-)
• His, pro, thr, leu, and F(-)
Transduction in Bacteria

• Transfer of bacterial DNA

transferred via
bacteriophage
• Bacteriophage
• Virus that infects bacteria
Transduction

• Steps of transduction
• 1- bacteriophage infects donor bacterial cell
• 2- Phage DNA and proteins, and bacterial chromosome is
broken into pieces
Transduction

• Steps of transduction
• 3- during phage reassembly, bacterial DNA incorporated in
capsid of bacteriophage
• 4 – donor cell lysis releasing new bacteriophage particles
Transduction

• Steps in transduction
• 5- phage carrying donor DNA infects new recipient cell
• 6- recombination can occur
• Producing bacteria with genotype different than donor and recipient
Transduction

• Generalized transduction
• Previously explained
• Specialized transduction
• Only certain genes are transferred
• i.e. phage codes for toxins to be produced
• Cornybacterium diphtheriae – diphtheria toxin
• Streptococcus pyogenes – erythrogenic toxin
• Escherichia coli – Shiga toxin (hemorrhagic diarrhea)
Plasmids

• Plasmids
• Self replicating rings of DNA
• 1-5% size of chromosomal DNA
• Non – essential genes
• Conjugative plasmid
• F factor
• Dissimilation plasmids
• Code for enzymes to
breakdown unusual sugars and
hydrocarbons
• Help in survival of unusual
environments
Plasmids

• Other plasmids
• Toxins (Anthrax, tetanus, Staph)
• Bacterial attachment
• Bacteriocins
• Toxic proteins that kill other bacteria
• Resistance factors (R factors)
• Resistance to antibiotics, heavy metals, cellular toxins
Plasmids

• Resistance factors
• Two groups
• RTF – resistance transfer factor
• Includes genes for plasmid replication and conjugation
• r-determinant
• Resistance genes
• Codes for production of enzymes that inactivate drugs or toxic substances
• Bacteria can conjugate and transfer plasmids between
species
• Neisseria
• Penicillinase resists penicillin
R factor Plasmids
Transpoons

• Transpoons
• Small segments of DNA that move from one region to
another
• 700-40,000 base pairs
• Occur in all organisms
• Can insert within genes
• Disrupt transcription of gene
• Occurs rarely (similar to spontaneous mutation rate)
Transpoons

• Transpoons
• Contain gene for transposition
• Insertion sequence (IS)
• Codes for transposase
• Cuts and seals DNA for transpoons
Review