MICROBIAL GENETICS

PART 1

Ø Genetic Material
Ø Central Dogma
Ø Genes Expression

TERMINOLOGY Levels of Genetic Study

Ø Genetics : the science of heredity
heredity;; the study of what
genes are, how they carry information, how
their information is expressed, and how
they are replicated and passed to
subsequent generations or other
organisms
Ø Heredity : the transmission of genetic information
from an organism to its off
off--springs
springs.. All
information necessary for life is stored in
an organism’s genetic material in DNA
(RNA in some viruses)
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Ø Genome : Sum total of genetic material of an organism
(chromosomes + mitochondria/chloroplasts and/or
plasmids) Ø DNA : A macromolecule composed of repeating units called
• genome of cells – DNA nucleotides. DNA exists as long strands of nucleotides
• genome of viruses – DNA or RNA twisted together to form a double helix. Each strand
Genome Size : has a backbone of alternating sugar and phosphate
• smallest virus – 4-5 genes groups and a nitrogenous base is attached to each
• E. coli – single chromosome containing 4,288 genes; 1 mm; 1,000X
longer than cell
sugar. Two strands are held together by hydrogen
• Human cell – 46 chromosomes containing 31,000 genes; 6 feet; bonds between the nitrogenous bases
180,000X longer than cell
Ø Nucleotide : Each nucleotide has three parts
Ø Chromosomes : Typically a circular (in Prokaryotes) or linear
l Nitrogenous Base; Adenine, Cytosine, Guanine, Thymine
(in Eukaryotes) thread like molecule of DNA.
l Deoxyribose (Pentose sugar ribose with one less oxygen)
Chromosomes contain the genes.
l Phosphate Group
Ø Genes : Segments of DNA (RNA in some viruses) that code for
functional products. Gene is basic unit of heredity

DNA STRUCTURE

Ø 2 strands twisted into a helix (double helix)

Ø Sugar - phosphate backbone
Ø Nitrogenous bases form steps in ladder
ALL GENES = DNA ?
l constancy of base pairing
or
l A binds to T with 2 hydrogen bonds
ALL DNA = GENES ?
l G binds to C with 3 hydrogen bonds

Ø Antiparallel strands 3’ to 5’ and 5’ to 3’

Ø One strand is the complement of the other
Ø Order of bases constitutes the DNA code

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Ø Complementarity of Base Pairing : The base pairs
always occur in a specific way
l Adenine always pairs with Thymine (in DNA)

l Cytosine always pairs with Guanine

• Because of this, complementary sequence of

bases on one DNA strand determines the
sequence of bases on other DNA strand
• The complementary sequence allows for the
precise duplication of DNA during cell division

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 Different Kinds of RNA
PLASMIDS
• rRNA • Combines with certain proteins to make
(Ribosomal RNA) ribosomes
• Provides a site for protein synthesis Ø Plasmids are self self--replicating circular extra
extra--
Ø Copy of DNA information for synthesis of
chromosomal molecules of DNA carrying
Ø mRNA
(Messenger proteins genes that are not usually essential for the
RNA) Ø Corresponds to one or more genes in DNA cell’s survival
survival..
Ø Have codons (triplet bases) Ø 1-5% of the size of bacterial chromosomes
Ø One or more ribosome can attach at a time Ø Found mainly in bacteria and some
* tRNA * Pick up amino acids in cytoplasm and transfer eukaryotes such as Saccharomyces
(Transfer RNA) them to mRNA cerevisiae (a yeast)
* Cloverleaf shape with attachment site for a
specific amino acid
* Has anticodon (triplet bases) complementary
to the codon in mRNA

PLASMIDS
TYPES OF PLASMIDS
1. Conjugative Plasmids
A prokaryotic plasmid that carries genes for sex
pili and for transfer of plasmid to another cell e.g.,
F factor plasmid
2. Dissimilation Plasmids
May code for enzymes that trigger the catabolism
of certain unusual sugars and hydrocarbon e.g.,
some Pseudomonas can use hydrocarbons of
petroleum as their primary carbon and energy
source
PLASMIDS
3.Plasmids carrying genes for toxins or bacteriocins
These enhance the pathogenicity of a bacterium
e.g., E. coli that cause infant diarrhea and
traveler's diarrhea carry this plasmid for toxin
production and attachment to intestinal wall
(without this plasmid, it is nonpathogenic)
nonpathogenic)..
Bacteriocins are proteins toxic for other bacteria
4. Resistance factor (R factor) plasmids
Carry genes that cause resistance to antibiotics or
cellular toxins
toxins..
Ø Particular plasmids can be transferred between
different species like Escherichia, Klebsiela
Klebsiela,, and
Salmonella

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 Transfer of Information C Flow of Genetic Information
E
DNA N
T
Replication R
DNA A
Reverse L
Transcription Transcription
D
RNA
O
G
Translation M
PROTEIN A

DNA REPLICATION
Ø The process of DNA replication is
Ø It takes place before every cell division so that each Semiconservative as each double stranded DNA
daughter cell get a copy of the complete genetic molecule has one original mother strand and one
material newly synthesized strand

Ø Two strands of the double helix separate to form a Ø DNA Polymerase enzyme can add new nucleotides
Replication Fork (the point at which replication occurs) at the 3`end of the newly synthesized strand only
only;;
so the DNA is synthesized in one direction
designated as
Ø Each strand then serves as a template for the
synthesis of a new strand following the rules of
complementary base pairing Ø 5` 3`only

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 R BACTERIAL REPLICATION
E
P
L
I
C
A
T
I
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DNA REPLICATION DNA REPLICATION

Ø Both strands of DNA are antiparallel (oriented in opposite

direction relative to each other)
Ø To add a new nucleotide to the growing strand of
3` 5` (Template) DNA, a nucleoside triphosphate bonds to the sugar
5` 3` (leading strand) and loses two of its phosphates
phosphates.. Hydrolysis of the
phosphate bonds provide energy for the reaction
reaction..
Ø At the replication fork, leading strand is synthesized
continuously as DNA Polymerase enzyme moves towards the
replication fork Ø In some bacteria like E. coli
coli,, two replication forks
move in opposite direction, so the replication is
Ø Lagging strand synthesis is discontinuous as it is made in Bidirectional.. Two forks ultimately meet each other
Bidirectional
fragments (called Okazaki fragments
fragments)) and the DNA Polymerase and the replication is complete
enzymes move in opposite direction of the growth of replication
fork

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 DNA REPLICATION (Steps)
Ø Helicase enzyme opens the double helix up, so a
replication fork is made (some other proteins are also
involved)
Ø RNA Polymerase makes a short piece of RNA (called
RNA Primer
Primer)) on the DNA template
Ø DNA Polymerase III then adds nucleotides at the 3 `
end of the RNA Primer & make the daughter strand
strand.. It
also does the proof reading to avoid mistakes in base
pairing
Ø DNA Polymerase I removes RNA Primer and
replaces it with DNA nucleotides but cannot connect
the two newly made segments together
Ø DNA Ligase joins the newly made DNA fragments
Ø DNA Gyrase creates twists or supercoils of DNA

GENES EXPRESSION TRANSCRIPTION

Ø It is the synthesis of complementary strand of RNA
Ø Genotype : from a specific gene (DNA template)
template).. Only one of the
The entire DNA or collection of genes
genes.. It is the entire two strands of DNA serve as template
genetic material of an organism
organism.. It represents the Ø There are three kinds of RNAs in bacteria
POTENTIAL properties l mRNA, tRNA
tRNA,, rRNA
Ø Transcription requires
Ø Phenotype : l RNA polymerase
l A supply of RNA nucleotides
It refers to the actual, EXPRESSED properties of an
Ø RNA Polymerase binds only to the Promoter Site on
organism.. It is an organism’s collection of proteins
organism the DNA
Ø RNA is synthesized in the 5` 3` direction too
Ø RNA synthesis terminates when RNA Polymerase
reaches a site on the DNA called the Terminator Site

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 TRANSLATION
Ø Protein synthesis is called translation because it involves
decoding the “language” of nucleic acids and converting
that information into the “language” of proteins
proteins..
Ø Language of mRNA is in the form of Codon (groups of
three nucleotides e.g., UCC AUC)
Ø Each codon codes for a particular amino acid (code is
called the genetic code)
Ø Initiator codon is AUG
Ø Terminator (stop or nonsense) codons
codons;; UAA, UGA, UAG
Ø Translation machinery includes :
l mRNA
l tRNA (which has anticodon
anticodon)) carrying amino acids
l Ribosome (have rRNA and specific proteins)

** Ribosomes moves along the mRNA in the 5` 3`

direction

STEPS INVOLVED IN TRANSLATION

Genetic code

Degeneracy of genetic
code

Necessary components assemble. Initiator codon (AUG) is the

point where translation begins

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 The first tRNA binds to the start codon, bringing the amino The first amino acid is transferred by the ribosome to the
acid methionine. Second tRNA arrives on second codon (it Second tRNA (peptide bond is formed)
must have complementary anticodon to the second codon)

Empty tRNA leaves. Ribosome then moves along the mRNA As the proper amino acids are brought into line one by one,
to the next codon toward 3` direction. New tRNA arrives with peptide bonds are formed between them, and a polypeptide
next amino acid chain is produced. Translation end at one of the stop codons

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STOP CODON (UAA, UAG, or UGA) ON m RNA SEPARATES THE PROKARYOTIC TRANSLATION
SYNTHETIC MACHINERY

mRNA Processing Before Translation In Eukaryotes

Differences between Eukaryote and Prokaryote
EUKARYOTE PROKARYOTE
Chromosome > 1, Linear 1, Circular
mRNA Monocistronic Polycistronic
DNA • Contains large regions of • Rarely contains any
repetitive DNA "extra" DNA
• Much of eukaryotic DNA • Often more than 95% of
does not code for proteins the genome codes for
proteins

Genes Split into exons and introns Almost never split

Transcription & Do not occur simultaneously Can take place
Translation simultaneously
Post- √ -
transcriptional
process
Start signal for Methionine Formyl-methionine
protein synthesis

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 PCR Animation
Please click here.
BIOTECHNOLOGY
Process

Denature

REPLICATION → DNA AMPLIFYING

Anneal Primer

POLYMERASE CHAIN REACTION

(PCR)
Replicate
DNA

1st cycle 2nd cycle 3rd cycle

MICROBIAL GENETIC PART 2

• REGULATION OF GENE EXPRESSION

• MUTATION
• GENE TRANSFER

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