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Chapter 9 - Microbial Genetics

Topics - Genetics - Flow of Genetics - Regulation - Mutation - Recombination


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Genetics
Genome (The sum total of genetic material of a cell is referred
to as the genome.)

Chromosome Gene Protein

Genotype Phenotype
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Chromosome
Procaryotic
Histonelike proteins condense DNA

Genes
Three categories
Structural Regulatory Encode for RNA

Eucaryotic
Histone proteins condense DNA

Subdivided into basic informational packets called genes

Genotype

Phenotype
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sum of all gene types Expression of the genotypes


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Representation of the flow of genetic information.

Flow of Genetics
DNA =>RNA=>Protein
Replication Transcription Translation

Fig. 9.9 Summary of the flow of genetic information in cell.

DNA
Structure Replication

DNA is lengthy and occupies a small part of the cell by coiling up into a smaller package.

Fig. 9.3 An Escherichia coli cell disrupted to release its DNA molecule.

Structure
Nucleotide
Phosphate Deoxyribose sugar Nitrogenous bases (purines- adenine, guanine;
pyramidines thymine, cytosine)

Purines and pyrimidines pair (A-T or G-C) and the sugars (backbone) are linked by a phosphate.

Double stranded helix


Antiparallel arrangement
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Fig. 9.4 Three views of DNA structure

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Replication
Semiconservative Enzymes Leading strand Lagging strand
Okazaki fragments

Semiconservative
New strands are synthesized in 5 to 3 direction

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Semiconservative replication of DNA synthesizes a new strand of DNA from a template strand.

Enzymes
Helicase DNA polymerase III Primase DNA polymerase I Ligase Gyrase
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Fig. 9.5 Simplified steps to show the semiconservative replication of DNA

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The function of important enzymes involved in DNA replication.

Leading strand
RNA primer initiates the 5 to 3 synthesis of DNA in continuous manner

Table 9.1 Some enzymes involved in DNA replication

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The steps associated with the DNA replication process.

Lagging strand
Multiple Okazaki fragments are synthesized Okazaki fragments are ligated together to form one continuous strand
Fig. 9.6 The bacterial replicon: a model for DNA Synthesis

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Replication processes from other biological systems (plasmids, viruses) involve a rolling cycle.

RNA
Transcription
Message RNA (mRNA) Transfer RNA (tRNA) Ribosomal RNA (rRNA)

Codon

Fig. 9.8 Simplified model of rolling circle DNA Replication

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Transcription
A single strand of RNA is transcribed from a template strand of DNA RNA polymerase catalyzes the reaction Synthesis in 5 to 3 direction

mRNA
Copy of a structural gene or genes of DNA
Can encode for multiple proteins on one message

Thymidine is replaced by uracil The message contains a codon (three bases)


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The synthesis of mRNA from DNA.

tRNA
Copy of specific regions of DNA Complimentary sequences form hairpin loops
Amino acid attachment site Anticodon

Participates in translation (protein synthesis)


Fig. 9.12 The major events in mRNA synthesis
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Important structural characteristics for tRNA and mRNA.

rRNA
Consist of two subunits (70S) A subunit is composed of rRNA and protein Participates in translation

Fig. 9.11 Characteristics of transfer and message RNA

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Ribosomes bind to the mRNA, enabling tRNAs to bind, followed by protein synthesis.

Codons
Triplet code that specifies a given amino acid Multiple codes for one amino acid 20 amino acids Start codon Stop codons

Fig. 9.9 Summary of the flow of genetics

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The codons from mRNA specify a given amino acid.

Representation of the codons and their corresponding amino acids.

Fig. 9.14 The Genetic code

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Fig. 9.15 Interpreting the DNA code

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Participants involved in the translation process.

Protein
Translation
Protein synthesis have the following participants
mRNA tRNA with attached amino acid Ribosome

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Fig. 9.13 The players in translation

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The process of translation.

Translation
Ribosomes bind mRNA near the start codon (ex. AUG) tRNA anticodon with attached amino acid binds to the start codon Ribosomes move to the next codon, allowing a new tRNA to bind and add another amino acid Series of amino acids form peptide bonds Stop codon terminates translation
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Fig. 9.16 The events in protein synthesis

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For procaryotes, translation can occur at multiple sites on the mRNA while the message is still being transcribed.

Transcription and translation in eucaryotes


Similar to procaryotes except
AUG encodes for a different form of methionine mRNA code for one protein Transcription and translation are not simultaneous Pre-mRNA
Introns Exons

Fig. 9.17 Speeding up the protein assembly line in bacteria

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The processing of pre-mRNA into mRNA involves the removal of introns.

Regulation
Lactose operon
sugar

Repressible operon
Amino acids, nucleotides

Antimicrobials

Fig. 9.18 The split gene of eucaryotes

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The regulation of sugar metabolism such as lactose involves repression in the absence of lactose, and induction when lactose is present.

The regulation of amino acids such as arginine involves repression when arginine accumulates, and no repression when arginine is being used.

Fig. 9.19 The lactose operon in bacteria

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Fig. 9.20 Repressible operon

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Antimicrobials
Ex. Antibiotics and drugs can inhibit the enzymes involved in transcription and translation

Mutations
Changes made to the DNA
Spontaneous random change Induced chemical, radiation. Point change a single base Nonsense change a normal codon into a stop codon Back-mutation mutation is reversed Frameshift reading frame of the mRNA changes

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Examples of chemical and radioactive mutagens, and their effects.

Repair of mutations involves enzymes recognizing, removing, and replacing the bases.

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Table 9.3 Selected mutagenic agents and their effects

Fig. 9.22 Excision repair of mutation by enzymes

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The Ames test is used to screen environmental and dietary chemicals for mutagenicity and carcinogenicity without using animal studies.

Effects of mutations
Positive effects for the cell
Allow cells to adapt

Negative effects for the cell


Loss of function Cells cannot survive

Fig. 9.23 The Ames test.

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Recombination
Sharing or recombining parts of their genome
Conjugation Transformation Transduction

Conjugation
Transfer of plasmid DNA from a F+ (F factor) cell to a F- cell An F+ bacterium possesses a pilus Pilus attaches to the recipient cell and creates pore for the transfer DNA High frequency recombination (Hfr) donors contain the F factor in the chromosome
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Conjugation is the genetic transmission through direct contact between cells.

Transformation
Nonspecific acceptance of free DNA by the cell (ex. DNA fragments, plasmids) DNA can be inserted into the chromosome Competent cells readily accept DNA

Fig. 9.24 Conjugation: genetic transmission through direct contact

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DNA released from a killed cell can be accepted by a live competent cell, expressing a new phenotype.

Transduction
Bacteriophage infect host cells Serve as the carrier of DNA from a donor cell to a recipient cell
Generalized Specialized

Fig. 9.25 Griffiths classic experiment in transformation

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Genetic transfer based on generalized transduction.

Genetic transfer based on specialized transduction.

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Fig. 9.26 Generalized transduction

Fig. 9.27 Specialized transduction

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Transposon
Jumping genes Exist in plasmids and chromosomes Contains genes that encode for enzymes that remove and reintegrate the transposon Small transposons are called insertion elements
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Movement of transposons can occur in plasmids and chromosomes.

Fig. 9.28 Transposons: shifting segments of the genome

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