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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
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
Induced mutations
Physical agents :
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
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
Figure 6.9
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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
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)
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
Replicative Transposition
Involves replication of the TE and insertion of the copy into another chromosomal location
Only found in bacteria
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
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
RECOMBINATION
classic recombination
common in DNA viruses
Reassortment
form of recombination (non classical)
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!
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
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