Microbial Genetics

Structure of Genetic Material

DNA & RNA

DNA=deoxyribonucleic acid
RNA=ribonucleic acid
Basic building blocks:
Nucleotides
Phosphate group
Pentose sugar
Nitrogenous base
 Structure of DNA
• Double stranded (double helix)
• Chains of nucleotides
• 5’ to 3’ (strands are anti-
parallel)
• Complimentary base pairing
• A-T
• G-C
 Bacterial DNA
• Bacteria have closed, circular DNA
• Genome: genetic material in an organism
• E. coli
• 4 million base pairs
• 1 mm long (over 1000 times larger that actual
bacterial cell)
• DNA takes up around 10% of cell volume
DNA Replication-occurs at the replication
fork

• 5’ to 3 ‘
• DNA helicase-unzips + parental DNA strand that is
used as a template
• Leading stand (5’ to 3’- continuous)
*DNA polymerase-joins growing DNA strand after
nucleotides are aligned (complimentary)
• Lagging strand (5’ to 3’- not continuous)
*RNA polymerase (makes short RNA primer)
*DNA polymerase (extends RNA primer then digests RNA primer
and replaces it with DNA)
*DNA ligase (seals Okazaki fragments-the newly formed DNA
fragments)
Replication Fork
Protein Synthesis
•DNA------- mRNA------ protein
transcription translation

Central Dogma
of Molecular Genetics
 Transcription
• One strand of DNA used as a template to make a
complimentary strand of mRNA
• Promoter/RNA polymerase/termination site/5’ to 3’
• Ways in which RNA & DNA differ:
• RNA is ss
• RNA sugar is ribose
• Base pairing-A-U
Transcription
 Types of RNA
• Three types:
• mRNA: messenger RNA
• Contains 3 bases ( codon)
• rRNA: ribosomal RNA
• Comprises the 70 S ribosome
• tRNA: transfer RNA
• Transfers amino acids to ribosomes for protein synthesis
• Contains the anticodon (3 base sequence that is complimentary to
codon on mRNA)
Genetic Code

• DNA: triplet code

• mRNA: codon (complimentary to triplet code of DNA)

• tRNA: anticodon (complimentary to codon)

Genetic Code

• Codons: code for the production of a specific amino acid

• 20 amino acids
• 3 base code
• Degenerative: more than 1 codon codes for an amino acid
• Universal: in all living organisms
Genetic Code
 Translation
• Three parts:
• Initiation-start codon (AUG)
• Elongation-ribosome moves along mRNA
• Termination: stop codon reached/polypeptide released
and new protein forms
• rRNA=subunits that form the 70 S ribosomes
(protein synthesis occurs here)
• tRNA=transfers amino acids to ribosomes for
protein synthesis)
Mutations
•Changes in base sequence of DNA/lethal
and inheritable
•Can be:
• Harmful
• Lethal
• Helpful
• Silent
Normal DNA/Missense Mutation
Missense-just one different amino acid formed-caused from a
base substitution, single base is replaced with a different one
Nonsense Mutation/Frameshift Mutation
Nonsense mutation-base substitution in the middle of the mRNA
results in the formation of the stop codon/protein synthesis stopped
Frameshift-1 or a few nucleotides are deleted or inserted-alters
3by3 transitional reading frame/produces inactive protein Base
substitutions and frameshift mutations occur spontaneously or by
chemicals in the environment
 Genetic Transfer in Bacteria
• Genetic transfer-results in genetic variation
• Genetic variation-needed for evolution
• Three ways:
• Transformation: genes transferred from one bacterium
to another as “naked” DNA
• Conjugation: plasmids transferred 1 bacteria to another
via a pilus
• Transduction: DNA transferred from 1 bacteria to
another by a virus
Transduction by a Bacteriophage
The mechanism of
genetic
transformation in
bacteria
Griffith’s experiment demonstrating
genetic transformation
Conjugation in E. coli
Conjugation in E. coli
Conjugation continued…
Conjugation continued…
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