L4: Microbial Genetics and Drug Resistance Flashcards
What is the definition of the genome?
It is the total genetic information in an organism.
What are the characters of the prokaryotic genome?
ο Consists of a single copy (Haploid) circular DNA molecule.
ο Range from 580-4600 Kbp
ο Many bacteria contain extrachromosomal DNA materials as a part of the genome called plasmids and transposons.
What is the definition of bacterial extrachromosomal elements?
- These are the DNA material present in a cell other than chromosomal DNA
- The most famous extrachromosomal DNA are:
A. Plasmid.
B. Transposons.
C. Bacteriophage (virus infecting bacteria)
What is the definition of plasmids?
Plasmids are pieces of DNA that exist separate from the chromosome, they contain an origin of replication so they replicate independently.
What are plasmids classified according to?
1- According to the size of the plasmid 2- According to copy number 3- Shape of plasmid 4- Moving plasmid from cell to cell 5- Artificial and natural plasmids 6- according to host range 7- according to compatibility
What are plasmids classified according to size?
Starting from a few hundred base pairs up to 3000 Kbp.
What are plasmids classified into according to copy number per cell?
1- Stringent plasmids
ο 1-2 copies / cell
ο like F- plasmid and phage β plasmid
hybrid (P1)
2- Low copy number plasmids
ο 10-15/copies/cell
ο such as pSC 101
3- High copy number plasmid
ο up to 50 copies/cell
ο like ColE, plasmid
4- Extremely high copy number plasmid
ο these are specifically engineered to be up to 100-200 copies/cell
What are plasmids classified into according to the compatibility of plasmids?
Compatible plasmids:
β’ a cell can maintain more than one plasmid in the same cell (if they carry
different origins of replication).
Incompatible plasmids:
β’ The inability of two plasmids to be maintained in the same cell (if they carry the same origin of replication
What are plasmids classified into according to the shape?
The shape of plasmids can be classified into 3 groups
1- Covalently closed circular (CCC) form:
ο The most common form
ο Present as ds completely closed circular forms (as in E.coli).
2- Semicircular form:
ο Transient form
ο Present as one strand is completely closed, the other strand is opened.
3- Linear:
ο unstable because it is attacked by exonucleases.
What are plasmids classified into according to host range?
1- Broad host range plasmids:
β’ Can replicate in a wide range of bacteria.
2- Narrow host range plasmids:
β’ only replicate in one or a few closely related bacteria.
What are plasmids classified into according to moving from one cell to another?
Conjugative plasmids: which have the tra genes that can mobilize the plasmid from one cell to another by conjugation
Shuttle vectors: plasmids that propagate in 2 diff. Hosts species (yeast and bacteria)
Nonβconjugative plasmids: Cannot be mobilized under any known conditions
What are plasmids classified into according to naturallity or artificiality?
1- natural plasmids:
β’ All the above plasmids are present naturally in bacterial and some yeast cells.
2- Artificial plasmids :
β’ are naturally present plasmid but designed artificially
β’ to be used in genetic cloning as vectors
β’ by adding antibiotic-resistant markers or DNA sequences to be the target of
restriction endonucleases.
What is the importance of plasmids?
- Resistance:
ο Antibiotic resistance.
ο Heavy metals (metal reductase).
ο U/V (DNA repair enzymes). - Conjugation.
- Production: Toxins & enzymes and bacteriocin.
- Biochemical reactions: Sugar fermentation.
- Molecular biology: As a vector.
ο Cloning vectors
ο Gene therapy: These are plasmids used for the insertion of therapeutic genes to express the protein that is lacking in the cells
What are transposons?
Extra-chromosomal small pieces of DNA those are capable of moving themselves from one location in DNA to another, (movable elements or jumping genes).
What are the 3 forms of transposons?
(a) Insertion sequence (IS)
(b) Composite transposons (Tn)
(C) Non β composite transposons
What are the characters of the insertion sequence?
ο The simplest form.
ο They encode only proteins needed for their own transposition
ο Carry repeated nucleotides at their ends (direct repeats or inverted repeats ~ (15-25).
ο Examples: IS 1, 3, and 10.
What are the characters of composite transposons?
Contain 2 IS at both ends and central piece of DNA which encode for antibiotic resistance, virulence factors.