Plasmids

Schematic drawing of a bacterium

with its plasmids.
(1) Chromosomal DNA. (2) Plasmids

RB Shahi
 Capable of autonomous replication
 i.e. self- replicating

 A plasmid is a DNA molecule separate from the

chromosomal DNA
 i.e.. Extra chromosomal(replicons)

 That are commonly present in bacteria, archaea, and yeasts

 Typically circular and double-stranded but Linear

plasmids have been found in a variety of bacteria,e.g.
Streptomyces sp. and Borrelia burgdorferi

 Often carry genetic information that is useful only under

atypical conditions, and are sometimes lost from the cell
(in non-selective environment)
 1952: J. Lederberg reviews the literature on cell
heredity and suggests the term "Plasmid" for all
extrachromosomal hereditary determinants.

 1954: Pierre Fredéricq and colleagues show that

colicine (plasmids) (large toxin proteins (50-70kD) )
behave as genetic factors independent of the
chromosome.

 1958: François Jacob and Elie Wollman propose the

term "Episome" to describe genetic elements such as F
factor, colicine, and phage lambda, which can exist
both in association with the chromosome and
independent of it.
 By the end of the 1960s, both the genetic and physical
nature of plasmids and cytoplasmic heredity had been
known in detail and the "Modern Period" of Plasmid
Research starts - recombinant DNA technology.

 1970s-80s: the Cytoplasmic mitochondrial and

chloroplast DNAs in green algae and plants were
continuously being studied and their circular forms of
dsDNAs.
 Plasmids frequently carry genes
associated
 Resistance to antibiotics, metal ions and
ultraviolet light;
 Virulence;
 Bacteriocin-production;
 Degradation of toxic organic compounds; and
 The ability to transfer from one bacterial cell to
another.
 Conformations of Plasmid DNAs
(Plasmid DNA may appear in the following five conformations)

1) "Supercoiled" (or "Covalently Closed-Circular") DNA is

fully intact with both strands uncut.

Super Coiled
2) "Relaxed Circular" DNA is fully intact, but "relaxed"
(supercoils removed). SC

3) "Supercoiled Denatured" DNA. small quantities occur

Relaxed region
following excessive alkaline lysis; both strands are uncut
but are not correctly paired, resulting in a compacted
plasmid form.

4) "Nicked Open-Circular" DNA

has one strand cut. Nicked DNAs

5) "Linearized" DNA has both Linear DNA

strands cut at only one site.
 Conformation Vs Electrophoretic mobility

The relative electrophoretic mobility (speed) of these DNA

conformations in a gel is as follows:

Nicked Open Circular

(slowest)

Linear

Relaxed Circular

Supercoiled Denatured

Supercoiled (fastest)
 Classification of Plasmids
• Based upon the number of copies per cell, plasmids are
classified into two types.
– Stringent plasmids
– Relaxed plasmids
• Based upon plasmid incompatibility: inability of two
different plasmids to co exist in the same cell in the
absence of selection pressure
– Compatible
– Incompatible
• Based upon the ability to take part in conjugation,
plasmids are of two types.
– Conjugative
– non conjugative
• Host Range
– Broad spectrum
– Narrow spectrum
• Phenotypic effects
– Fertility
– Bacteriocinogenic plasmid
– Resistance plasmid (R factors)
– Etc.
 Stringent plasmids
• These plasmids exist in small numbers, i.e.
<100 copies/cell (limited number of copies
per cell)
• Under the control of bacterial genome for
replication and segregation
• Generally, conjunctive plasmids are mostly
stringent plasmids.
 Relaxed plasmids
• These plasmids exist in large numbers, i.e.,
> 100 copies/cell.(multiple copies per Cell)
• not under the control of bacterial genome
for replication and segregation
• Generally they are of low molecular weight
• most of them are of the non conjugative
type.
 Compatible

• Have different replication control

 Incompatible
• Have same replication control
 Conjugative

• those which take part in conjugation

• They have tra genes which help in
conjugation,
• e.g. F. Plasmids
 non conjugative
• those which does nittake part in conjugation
• They lack tra genes which help in
conjugation,
• e.g. pBR 322
 Broad spectrum
• Replicate in many types of bacteria
– RSF 1010(wide range of Gram-negative and
Gram-positive bacteria-non conjugative)
– RP4(most Gram-negative bacteria-conjugation)
– etc
 Narrow spectrum

• Replicate in only one type of bacteria

– pBR322 in E .coli
 Maintenance of Plasmids
• Two factors are important
– One is their faithful replication at least once
in each cell cycle: Regulated
1. by antisense RNA and
2. by binding of essential proteins to iterons
– their accurate partition so that each daughter
cell receives at least one copy
Rop=Repressor of Primer
 Function of Plasmids
1) Antibiotic resistance
2) Antibiotic production
3) Degradation of aromatic compounds
4) Haemolysin production
5) Sugar fermentation
6) Enterotoxin production
7) Heavy-metal resistance
8) Bacteriocin production
9) Induction of plant tumours
10)Hydrogen sulphide production
11)Host-controlled restriction and modification
What are cryptic plasmids?
 Based on their function, there are
five main classes
 Fertility-(F)plasmids: they are capable of
conjugation or mating.
 Resistance-(R) plasmids: containing antibiotic or
drug resistant gene(s). Also known as R-factors,
before the nature of plasmids was understood
 Col-plasmids: contain genes that code for colicines,
proteins that can kill other bacteria.
 Degrative plasmids: enable digestion of unusual
substances, e.g., toluene or salicylic acid.
 Virulence plasmids: turn the bacterium into a
pathogen. Hly Plasmid :α-haemolysins of RBCs
Plasmids can belong to more than one of these functional groups.