Microbial Genetics

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Microbiology (Donato)
Marissa Alvarez
Flashcards by Marissa Alvarez, updated more than 1 year ago
Marissa Alvarez
Created by Marissa Alvarez almost 7 years ago
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Question Answer
Vertical Gene Transfer
Horizontal (Lateral) Gene Transfer
Characteristic of the Bacterial Genome -Population is CLONAL b/c bacteria REPLICATE by CELL DIVISION
How do bacteria introduce variety or diversity into their genome? (2 ways!) 1.) Mutation (from existing genetic info.) 2.) Exogenous genetic info. (incorporating foreign DNA via horizontal (lateral) gene transfer) *20-30% of total bacterial genome comes from exogenous genetic info.
Mechanisms for Acquiring Exogenous DNA (3 ways!) 1. Transformation 2. Transduction 3. Conjugation
1. TRANSFORMATION free or "naked" DNA transferred -DIRECT uptake of naked DNA -NO donor bacterium required -can happen two ways: with DNA fragments or with a PLASMID
Transformation with DNA Fragments: (Neisseria can do this naturally) With DNA fragments: 1. ds DNA binds to bacterial surface 2. nucleases degrade 1 strand 3. 1 DNA strand enters the cell 4. DNA integrates into chromosome by homologous recombination (or is degraded if unsuccessful) *Homologous recombination: a type of genetic recombination in which nucleotide sequences are exchanged between two similar or identical molecules of DNA
Transformation with PLASMIDS: With Plasmid: 1. plasmid binds to bacterial surface 2. WHOLE plasmid is taken up by bacterium with ease 3. Stable transformation
2. TRANSDUCTION -transfer of genetic information from donor to recipient by BACTERIOPHAGE -DNA transferred via phage (can be lytic or lysogenic) *Bacteriophage/phage = virus that infects bacteria
Bacteriophage or Phage CAPSID: head that surrounds and protects nucleic acid SHEATH: hollow cylinder used to inject phage genomic material TAIL FIBERS: used to attach to bacterial surface; determines SPECIFICITY
BASIC steps of the Lytic Cycle of Phage 1. Phage binds to bacterial 2. Injects viral nucleic acid (using the sheath) 3. Takes over host replication machinery for viral replication 4. Package/Assemble DNA into new phage heads 5. Lyse bacterial cell and release 100's of progeny (virions)
GENERALIZED TRANSDUCTION BACTERIAL DNA gets incorporated into phage head ACCIDENTALLY and is transferred to recipient bacterium (occasionally occurs) -happens as a result of error during construction of phage particles
Lysogeny (an alternative to lyses/lytic) Phage incorporate their DNA into the bacterial chromosome (becomes a prophage)
Plaque Assay Lytic phage LYSE bacteria growing in a lawn and produce a clearing that is visible and countable on an agar plate (creates plaques) Pour-plate technique: mix phage and bacteria together, add to soft agar, and overlay onto an agar plate -The SOFT agar (not as solid as normal agar plate) allows released phage progeny to migrate through it to infect subsequent bacterial cells, while the bacteria are prohibited from moving (due to difference in sizes between phage & bacteria)
Phage Therapy Prior to the advent of antibiotics, phage were used as antimicrobial agents Since then, this practice has stopped everywhere except in the former Soviet Bloc Today, with increasing antibiotic resistance among bacteria, the concept of “phage therapy” is being revisited Although NOT approved for HUMAN infections, phage therapy is being employed in the U.S. for FOOD safety EX: (Intralytix; Listeria, EHEC & Salmonella
3. CONJUGATION "Bacterial SEX" **requires DIRECT bacterium-to-bacterium contact -differs in Gram + and Gram - bacteria -mediated by conjugative PLASMID
Gram Positive (+) Bacterial Conjugation G+ form a MATING BRIDGE through SURFACE ADHESIONS
Gram Negative (-) Bacteria G- have a sex (F) plus for INTIMATE contact
CONJUGATION mediated by? example? HFr? Mediated by self-transmissible plasmid= CONJUGATIVE PLASMID, Example: F PLASMID F plasmid encodes for F pilus and genes -needed for transfer of DNA -Gaining the F plasmid = F+ If F plasmid incorporates into the chromosome = Hfr (high frequency recombinant), which can then transfer even larger pieces of DNA to recipient
F+ has the F gene (F plasmid that encodes for F plus and transfer genes) can donate ONLY to F-
F- recipient for F+ -will become F+ after receiving the F plasmid from an F+ bacterium
Hfr (high frequency recombinant) Will the F- likely remain F- or become F+ after conjugation with an F+ HFr cell? It is more likely to remain F- because an incomplete F plasmid would be transferred. (RARE for the whole plasmid to be transferred and for that cell to become a F+ HFr cell)
GENOMIC ISLANDS LARGE, discrete DNA segments differing b/w closely related bacterial strains (NON-CONSERVED) Acquired via HORIZONTAL GENE TRANSFER Recognizable by a SHIFT in GC-content Harbor sequences involved in GENETIC MOBILITY (e.g. transposases, integrases etc.) Often carry genes that offer a SELECTIVE ADVANTAGE, a benefit (e.g. antibiotic resistance, virulence-related genes etc.) -Can have Pathogenicity islands
Pathogenicity islands genomic islands that carry genes associated with causing disease (virulence associated genes) MOST important: encodes for everything the bacterium needs to be PATHOGENIC i.e., secretion systems, toxins, adhesins, invasins etc...
CONCLUSION *The NET effect of HORIZONTAL GENE TRANSFER (in addition to mutation & mobile elements) is the INCREASE in the genome's plasticity: addition of EXOGENOUS DNA contributes to bacterial DIVERSIFICATION
SIGNIFICANCE Bacterial genome= core genome (70-80%) + flexible gene pool (20-30%) Horizontal gene transfer leads to the introduction & stable inheritance of new genes May acquire virulence factors, antibiotic-resistance genes etc.- make a “better” pathogen, alter niche, evade host response Increase gene repertoire Contributes to diversification & adaptation of bacteria Provides genetic tools for modern day biotechnology & genetic engineering
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