Vous êtes sur la page 1sur 52

Bacterial and Viral Genetics

Dr. Shobha Broor Professor Dept. of Microbiology

What is genetics
Genetics study of heredity and variation Unit of heredity gene Gene : segment of DNA carrying information for a specific polypeptide Codon : sequence of 3 nucleotides coding for a single amino acid

Bacterial genetics
Genetic information in bacteria is coded by chromosomal DNA 3 types of RNA in bacteria:
m-RNA t-RNA ribosomal-RNA

Bacteria also contain extra chromosomal elements i.e.:


Plasmids or episomes (integrated with chromosomal DNA

Plasmids
Circular DNA molecules which can replicate independent of host genome Not essential for function of host bacteria but carry certain supplementary important genetic information for properties like :
Drug resistance Toxin production

Variations in bacteria
Phenotypic : Expression of genes in a particular environment Genotypic:
Mutations Gene transfer

Mutations
Heritable genetic changes which are random Frequency of spont. mutations ranges from 102-106 /per division Mutations can be
Spontaneous Induced

Mutations caused by
Base substitution Deletion Insertion rearrangements

Mutations may be point mutations or multisite mutations

Substitution :arise by mispairing


Transition: Pyrimidine to pyrimidine (usually no change in codon) Tranversion : Pyrimidine to purine ( codon change) Missense mutation: No discernible phenotype effect

Deletions or Insertions cause frame shift: coding message read incorrectly

Induced mutations
Physical agents :

UV light Radiation eg. Xrays Visible light Heat


Nitrous acid Acridine dyes 2-aminopurine 5-bromouracil

Chemical agents :

Suppressor mutation
A mutation at a 2nd locus which restores the lost phenotype

Gene transfer
Conjugation
direct physical contact & exchange

Transduction
phage

Transformation
uptake from environment

Transformation
Bacteria take up extracellular DNA Discovered by Frederick Griffith,1928, while working with strains of Streptococcus pneumoniae There are two types
Natural transformation
DNA uptake occurs without outside help

Artificial transformation
DNA uptake occurs with the help of special techniques

Griffiths experiment 1928

S or smooth strain lethal to mice

R or rough strain does not kill mice

Griffiths experiment 1928

However, if he were to take a mixture of the heat-inactivated S strain, mixed with the R strain, the mice would die. Thus there was some Material in the heat-killed S strain that was responsible for "transforming" the R strain into a lethal form.

Transformation
Natural transformation occurs in a wide variety of bacteria Bacteria able to take up DNA = competent bacteria carry genes encoding competence factors
proteins that uptake DNA into bacterium & incorporate it into the chromosome

Transformation
Artificial or forced transformation By treatment of cells with CaCl2 and temp. shock Plasmids can be transformed in to well characterized bacteria

Transduction
Bacteriophage mediated transmission of a portion of DNA from one bacteria to another During assembly of bacteriophage it may incorporate accidentally host DNA, - packaging error Usually caused by temperate phages which cause lysogeny phage DNA integrated with bacterial chromosomal DNA e.g. Diphtheria tox gene If only a particular gene is transduced : Restrictive transduction

It will undergo the lytic cycle

Prophage can exist in a dormant state for a long time

Virulent phages only undergo a lytic cycle

Temperate phages can follow both cycles

Figure 6.9

Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display

6-32

Transduction

Any piece of bacterial DNA can be incorporated into the phage This type of transduction is termed generalized transduction

Conjugation
Plasmids are responsible for conjugation Transfer of genetic material between 2 bacteria by contact is called conjugation Some plasmids have trs gene which gives the property for transfer of DNA called F plasmids Bacteria carrying these plasmids called F+ or donor bacteria These F+ bacteria have sex pili which help in attachment of F+ bacteria to F- bacteria and a bridge between the two is formed

Conjugation

Conjugation

Conjugation
Results of conjugation
recipient cell acquires F factor converted from F to F+ cell

F factor plasmid may carry additional genes


called F factors

F factor transfer can introduce genes & alter recipients genotype

Hfr Conjugation
When it exists as a free plasmid, the F plasmid can only transfer itself. This isnt all that useful for genetics. However, sometimes the F plasmid can become incorporated into the bacterial chromosome, by a crossover between the F plasmid and the chromosome. The resulting bacterial cell is called an Hfr, which stands for High frequency of recombination. Hfr bacteria conjugate just like F+ do, but they drag a copy of the entire chromosome into the F- cell.

Hfr Strains
1950s, Luca Cavalli-Sforza discovered E. coli strain very efficient at transferring chromosomal genes
designated strain Hfr (high frequency of recombination)

Hfr strains result from integration of F' factor into chromosome

Conjugation
Conjugative plasmid DNA replicates and a copy passes from donor to recipient along the sex pili When F factor is integrated with host cell chromosomal DNA called as HfR cell as these bacteria transfer chromosomal genes at a high frequency to recipient cells Another conjugation plasmid is RTF or Resistance transfer factor RTF transfers drug resistance genes

Transposons
Transposable genetic elements Ability to move from one plasmid to another or from plasmid to chromosome Also called jumping genes Transfer of genetic material called transposition and does not require any DNA homology

Transpositions
3 general types of transposition
Simple transposition Replicative transposition Retrotransposition

Simple Transposition

Known as Insertion Sequences - IS bacterial


Tn10

Replicative Transposition

Involves replication of the TE and insertion of the copy into another chromosomal location
Only found in bacteria

Simple & Replicative Transposons


Both contain a gene encoding a transposase enzyme Transposase function
recognizes direct and indirect repeats cuts DNA for both excision and insertion

Regulatory Sequences of Transposable Elements

transposon

Direct repeats DNA sequences that are identical and run in the same direction (53)
5 ATGACTGAC 3 3 TACTGACTG 5
and

5 ATGACTGAC 3 3 TACTGACTG 5

Inverted repeats - DNA sequences that are identical (or very similar) but run in opposite directions
5 CTGACTCTT 3 3 GACTGAGAA 5
and

5 AAGAGTCAG 3 3 TTCTCAGTC 5

Viral genetics
DNA chromosomes of eukaryotic host organisms generally require geologic time spans to evolve to the degree that their RNA viruses can achieve in a single human generation.

VIRAL GENETICS
VIRUSES GROW RAPIDLY A SINGLE PARTICLE PRODUCES A LOT OF PROGENY DNA VIRUSES SEEM TO HAVE ACCESS TO PROOF READING, RNA VIRUSES DO NOT SEEM TO

NATURE OF GENOMES
RNA or DNA

SEGMENTED OR NON-SEGMENTED

Viral genetics
Viruses undergo much more heritable changes as compared to bacteria as in RNA viruses RNA polymerase does not have proof reading capacity Genetic changes
Mutations Re-combinations and re-assortments

ORIGIN OF MUTATIONS
SPONTANEOUS
tautomeric form of bases polymerase errors
mutation rates usually higher in RNA viruses (lack of proof reading)

ORIGIN OF MUTATIONS
SPONTANEOUS PHYSICALLY INDUCED UV light , especially problem if no access to repair X-rays CHEMICALLY INDUCED

TYPES OF MUTATION
POINT

INSERTION DELETION

Phenotypic changes caused by Mutations


Conditional lethal mutations : mutations which are lethal to virus under one set of conditions termed non-permissible - wild-type (wt) grows under both sets of conditions
Temperature-sensitive (ts) mutants: do not grow at higher temperature (altered protein) Host-range mutants do not grow in all the cell types that the wt doesHost range mutants

PLAQUE SIZE
may show altered pathogenicity

DRUG RESISTANCE
important in the development of antiviral agents

ENZYME-DEFICIENT MUTANTS
some genes can be optional in certain circumstances

HOT MUTANTS
grow better at elevated temperature than wt less susceptible to host fever response

ATTENUATED MUTANTS
milder (or no) symptoms vaccine development pathogenesis

Genetic interactions among viruses


When 2 or more viruses infect the same host cell they may interact in a variety of ways :
Recombinations : Nucleic acid shows a break and a part of genome of one parent is joined to the genome of 2nd parent Reassortment : In segments virus genomes , a segment of genome of one virus is replaced with the segment from another virus

RECOMBINATION
classic recombination
common in DNA viruses

Reassortment
form of recombination (non classical)

very efficient segmented viruses only used in some new vaccines

Antigenic shift

Genetic reactivation
Special case of recombination or reassortment in which one or both the parental viruses are not infectious but in mixed infection , infectious progeny is produced that contain markers from both parents
Marker rescue: Occurs between genome of an active and inactive virus. A portion of genome of inactive virus are rescued by recombination and appear in viable progeny e.g. HDV

DEFECTIVE VIRUSES
lack gene(s) necessary for a complete infectious cycle helper virus provides missing functions
package me! copy me!

package me!

copy me!

DEFECTIVE INTERFERING (DI) VIRUSES (PARTICLES)


decrease replication of helper virus compete for viral precursors, etc. may modulate wt infections occur naturally eg. DI measles virus in subacute sclerosing panencephalitis SSPE

Multiplicity reactivation
When many virus particles interact in the same cell to generate a viable virus. Heavily damaged viral preparation used to infect cells at a high multiplicity of infection. Recombination occurs between damaged nucleic acids of the parents producing a viable genome which can replicate

COMPLEMENTATION
Interaction at the functional level, NOT the nucleic acid

level

ts mutant 1

ts mutant 2

Progeny virus assembled using wt N and wt M proteins Genomes in progeny are either ts M or ts N
mutants which can complement are generally in different genes

PHENOTYPIC MIXING

no changes in genome

possibly altered host range


possibly resistant to antibody neutralization Not a stable genetic change

Interference
Infection of a cell or an animal with two viruses often leads to inhibition of multiplication of one of the viruses , this effect is called Interference

Vous aimerez peut-être aussi