Withey: Bacterial Genetics Flashcards
Chromosome: Shape bps/gene #: Typical gene: Nucleus? Nucleoid def: Cs #:
Haploid and circular
~4,000,000 bp (~4,000 genes)
Typical Gene: 1000bp
No nucleus, introns or histones
Still highly structured, using histone-like proteins for form a nucleoid
Usually only have one chromosome
Plasmids and bacteriophages definition:
Plasmid: circular, extrachromosomal elements
Bacteriophage: bacterial viruses, integrated or autonomous
Plasmids:
Replication:
Size:
Quantity:
Episome:
Replication: autonomously replicating DNA (have their own origin of replication; can replicate independent of the chromosome)
Size: 5000-200,000 bps
Quantity: 1-500 per cell
Epsiome: a plasmid that can integrate into chromosome; some encode elements required for conjugation
Plasmid-Encoded Virulence Factors (5):
o Heat labile and heat stable toxins of E.coli
o Tetanus toxin of Clostridium tetani
o Anthrax toxin of Bacillus anthracis
o Shigella spp.’s ability to invade colonic epithelium
o Antibiotic resistance (in some circumstances)
Episomes/Resistance Factors:
- R Factors:
R Factors: conjugative episomes that encode antibiotic resistance
R Factors
Composed of 2 Subunits:
Composed of 2 Subunits:
Resistance Transfer Factor (RTF): allows for autonomous replication and conjugal transfer
Resistance Determinant: composed of one or more transposons, which carry the antibiotic resistance gene
- Transposons mediate the formation/resolution of R factors
Transposons (Tn)
Definition:
Composition
Definition: a sequence of DNA that can “hop” from place to place. An insertion sequence that has assimilated a drug-resistance gene.
Composition: antibiotic resistance gene flanked by insertion sequences, which encode for transposon mobility and allow for entry into host genome
Transposons (Tn)
Function:
Complex Transposons:
Function: disseminate antibiotic resistance
o Carried on a conjugative episome
o Hop into chromosome (overcome host restriction barriers)
Complex Transposons: consists of drug resistance (and other genes) flanked by 2 different insertion sequences
Transposons (Tn)
Example of Resistance:
Enterobacteriaceae have transferred ampicillin resistance to Haemophilus influenza and Neisseri gonorrhoeae
This concept of transferred resistance is the rationale behind using combinations of unrelated antibiotics
Bacteriophage definition:
Two types:
Bacteriophage: viruses that only infect bacteria
Two Types:
o Lytic Phages: infect, reproduce and kill bacteria by lysis
o Temperant Phages: integrate into chromosome to form lysogen or prophage
Examples of toxin/virulence factor genes that are carried in phage genomes:
How can bacteriophages be used as therapy?
Many toxin/virulence factor genes are carried in phage genomes:
o Examples: Vibrio cholera, E.coli
Can be used as therapy to kill antibiotic resistant bacteria
- Phage specific for a bacterial species can be isolated in a few days (very quick)
- They are very specific for their host bacterial species (protect normal flora)
- No effect on eukaryotic cells
Gene transfer restriction/modification:
A. Bacterial “immune system”
B. Defends against foreign DNA
C. Modification is the species-specific methylation of certain DNA sequences
D. Restriction is cleavage of unmethylated DNA at the same sequences by restriction enzymes
- Properly modified DNA is protected from cleavage by restriction enzyme
Transformation
Basics:
Competence definition:
Transformation is sensitive to what?
- Transformation:
- Basics: uptake of DNA from extracellular milieu (species-specific, sequence specific or non-specific)
Naked DNA adsorbs to bacteria and enters cytoplasm
Competence: ability to accept DNA; mechanisms vary among bacteria
Transformation is DNase sensitive
Transformation
Incoming DNA must recombine with host chromosome using:
Entry vs incorporation:
Incoming DNA subject to:
What must the incoming DNA have for RecA to function?
Incoming DNA must recombine with host chromosome using RecA enzyme
Any DNA may gain entry, however this does not mean it will be incorporated
Incoming DNA subject to host restriction barriers (Restriction/Modification system)
Incoming DNA must have some sequence homology with the host DNA for RecA to function
Transduction definition:
Transfer of genetic information by bacteriophage (phage)
Phage can be lytic (produce more phage, kill host cell) or lysogenic (integrate into host chromosome, do not kill host cell)
Generalized Transduction:
What is a pseudovirion?
Transferred DNA must:
Generalized Transduction: indiscriminate transfer of chromosomal sequences
Phage “accidentally” packages host sequences in pseudovirion (new phages made that have some host DNA)
Transferred DNA must integrate into recipient chromosome (RecA)
Specialized Transduction:
Can only occur via:
Specialized Transduction: transfer of specific chromosomal sequences
Can only occur through lysogenic phages
Specialized Transduction
Steps:
Bacteriophage specifically integrates into host chromosome (to form prophage)
a) Integrated prophage is lysogenic
b) Integration is site-specific & reversible
DNA damage induces excision of the bacteriophage
a) Pieces of chromosome pulled out with phage
b) Chromosome + phage DNA transferred to next host