Sub Topics:

A. DNA and Genetic Information

B. DNA Replication
C. Protein Synthesis:
Transcription and Translation
D.Gene Regulation and
Expression- Lac Operon
Introduction
 DNA as genetic material,
carry and pass the genetic
information from parents to
offspring.
 Genetic information – is
contained in the sequence of
nucleotide bases in DNA
molecule.
Describes the flow of genetic information from DNA
to RNA to Proteins.
•DNA Replication
•Transcription- is the synthesis of an RNA copy of a
segment of DNA.
•Translation
Structure of DNA
 Structure of DNA – Watson – Crick model.
 2 polynucleotide strands are linked together by
hydrogen bonds formed complementary bases.
 2 hydrogen bonds between adenine and thymine.
 3 hydrogen bonds between guanine and cytosine.
 DNA molecule showing anti-parallel 5’-3’ and 3’-5’
strands.
 Codon in DNA read from 3’-5’.
 2 nucleotides are joined together by covalent
phosphodiester bond to form dinucleotide.
Structure of RNA
 Single strand polynucleotide.
 3 types of RNA is mRNA, tRNA and rRNA.
mRNA
 Long single strands of RNA that are transcribed from
DNA.

 Codon in mRNA read from 5’to 3’.

 Contains information which allow cells to make

proteins.

 It will travel from nucleus to ribosomes and provide the

blueprints for all the proteins to be made by ribosomes.
rRNA
 RNA found in ribosomes

 Part of the structure of ribosomes

 During polypeptide synthesis, ribosomes provide the

site for protein synthesis.

 Ribosomal RNA provides a mechanism for decoding

mRNA into amino acids and interacts with tRNAs
during translation
tRNA
 Composed of around 80
nucleotides arranged in a
clover leaf form.

 Made in the nucleus and

then transported to the
cytoplasm.

 It picks up the necessary

amino acid in the cytoplasm
and carries to the surface of
ribosome for polypeptide
formation.
DNA as Carrier of Genetic Information
 Chromosomes known as the genetic material consist
of DNA, protein and RNA.

DNA
So which
one is the
material
genetic??? RNA

Protein
 Scientist found more RNA and protein in cytoplasm.

 Thus, possibility DNA is the genetic material.

 DNA content are same in different cells but not for

RNA and protein.

 Some experiments proved that DNA as the genetic

material:
 Griffith experiment in 1931.
 Avery et al. experiment in 1944.
 Hershey-Chase experiment in 1952.
Frederick Griffith (1931)
 Experiments with Streptococcus pneumoniae
 Virulent Strain (Strain-S)
 Forms a smooth colony.
 Cells secrete a polysaccharide capsule which are virulent and
pathogenic to most mammals and lead to death.
 Non-virulent Strain (Strain-R)
 Forms a rough colony.
 Cells do not secrete any capsule.
 Non-pathogenic
 Susceptible
 Both strain are very sensitive to heat.
Conclusion….
 This is cause by TRANSFORMING AGENT .

What is the transforming

agent?

 Transformation is a types of genetic transfer found in

bacteria.
 When heated to kill the S
strain, it also ruptured
the bacteria’s cell wall
and released the content.

One of the
component of S
strain was then
capable of Griffith found
transforming R the S strain in
strain into S strain the dead
when mix in the mouse.
same media.
Avery et al. Transformation Experiment (1944)
Oswald T. Avery

Colin MacLeod

Maclyn McCarty

 Avery repeated Griffith’s experiment by mixing the component

with R strain.
 Only the mixture containing nucleic acids and R strain
transformed to pathogenic S strain.
DNA as genetic material…
DNA as transforming agent
S strain R strain
(killed) + (living)

protease RNase DNase centrifugation

Protein RNA DNA Lipid

destroyed destroyed destroyed eliminated

Living Living Living Living

S cells S cells R cells S cells

No transformation
Oswald T. Avery’s Transformation Experiment - 1944

Determined that “IIIS” DNA was the genetic material

responsible for Griffith’s results (not RNA).

Peter J. Russell, iGenetics: Copyright © Pearson Education, Inc., publishing as Benjamin Cummings. 23
DNA as genetic material…
Avery et. al (1944)

conclusion
The strain without DNA caused no
transformation occur.

So, the transformation agent is DNA!

 Hershey-Chase experiment (1952)
• Bacteriophages = Virus that
attacks bacteria and replicates
by invading a living cell and
using the cell’s molecular
machinery.

Structure of T2 phage

• Bacteriophages are composed

of :
• Protein – makes up the
exterior (head and tail)
• DNA – located in head
Life cycle of virulent T2 phage:

26
 To determine that DNA was the genetic material of
the phage T2.
 They designed an experiment that could label
protein or DNA which entered the E. coli cell during
infection.
 Grew one batch of T2 phage in the presence of
radioactive sulfur, marking the proteins but not DNA.
 And grew another batch in the presence of radioactive
phosphorus, marking the DNA but not proteins.
 They allowed each batch to infect separate E.
coli cultures.
 They spun the cultured infected cells in a blender,
shaking loose any parts of the phage that remained
outside the bacteria.
 The mixtures were spun in a centrifuge which
separated the heavier bacterial cells in the pellet
from lighter free phages and parts of phage in the
liquid supernatant.
 They found that when the bacteria had been
infected with T2 phages that contained radio-
labeled proteins, most of the radioactivity was in
the supernatant, not in the pellet.
 When they examined the bacterial cultures with
T2 phage that had radio-labeled DNA, most of the
radioactivity was in the pellet with the bacteria.
 Hershey and Chase concluded that the injected
DNA of the phage provides the genetic
information that makes the infected cells produce
new viral DNA and proteins, which assemble into
new viruses.
Radioactive
isotope of DNA
– phosphorus,
Protein –
sulphur
 The One-Gene-One Enzyme
Hypothesis
 The theory that each gene is
responsible for the synthesis of a
single polypeptide.
 George Beadle and Edward Tatum
were among the first to investigate
biosynthetic pathways.
 They studied growth variants of
the fungus, Neurospora crassa.
 Their proposal, the one-gene-one
enzyme hypothesis came out of
their experiments.
 Beadle and Tatum’s
 Did exp. on Neurospora crassa (bread mold)
 Normal spores – germinate on minimal medium
 Mutant spores – unable to germinate
 Lost ability to synthesize non- essential amino acids i.e.
arginine
 Arginine biosynthetic pathway involves 4 different
enzymes located at 4 different loci on chromosome
 Each mutant had a single enzyme deficiency caused by
mutation at a single locus i.e. defect in one gene prevent
the synthesis of functional enzyme required to make a
particular amino acid.
Gene Hypothesis…
Neurospora crassa able to synthesize all of the
amino acids and other chemicals needed for
growth

Mutations will affect a single genes and single

enzymes in specific metabolic pathways.
Case 31
Case2 Gene

Enzyme

Amino acid

Wild type neurospora

Mutant neurospora

Minimal Nutrient Minimal Nutrient

 How to make sure ONE genes is only code ONE polypeptide???
Is the
Is the gene
gene AB
C only
only
Precursor produce enzyme
produce enzyme AB C only?
only?

X
Gene A X
Enzyme A

+
Ornithine
Ornithine

X
Gene B
X
Enzyme B

+
Citrulline
Citrulline

X
Gene C
X
Enzyme C

Arginine

+
Arginine
Gene Hypothesis…
Beadle & Tatum Experiment

conclusion
Each gene will produce certain enzyme
(polypeptide)

One gene one polypeptide.

DNA Replication Model
 Hypothesis 1
Conservative

Both parent strand

remain together or act
as template and all new
copies is made.
 …

Hypothesis 2
Semiconservative

The 2 strand of the

parental molecule
separate and each
functions as a template
for synthesis of a new
complementary strand
 Hypothesis 3
Dispersive

Each strand of both

daughter molecules
contains a mixture
of old and newly
synthesized parts
Process of copying a double stranded DNA strand
which is the two resulting double strands are
identical and each of them consist of one original
and one newly synthesize strand.

Proposed by Matthew
Meselson (left) and
Franklin W. Stahl (right)
in 1958.
  DNA has the ability to replicate.
 Basically what happen is:

DNA strand DNA strand Replication process

separated Replicated DNA
occurring
 Meselson & Stahl (1957)

Studied the replication

in E. coli

Uses 3 hypotheses

conservative semiconservative dispersive

Which one is the model of

DNA replication?
DNA Replication Model…
Meselson & Stahl (1958)
DNA Replication Model…

Result

First replication

Second replication
DNA Replication Model…

Result

First replication

Second replication
DNA Replication Model…

Result

First replication

Second replication
DNA Replication Model…
Meselson & Stahl (1957)

First
replication

Second
replication
DNA Replication Model…
Meselson & Stahl (1957)

DNA replication is take place by

semiconservative.
DNA Replication…
➢DNA Replication is semi-conservative.

➢Replication begins at special sites called origin of

replication.

➢A stretch DNA replication have a specific sequence of

nucleotides.

➢Protein that initiate DNA replication recognize this

sequence and attach to the DNA.

➢Two strands of DNA separate and opening up the

replication bubble.
 Steps in replication:
1. Initiation by replicator
initiation proteins at AT-rich
region
2. Unwinding of DNA molecule
making 2 separate template
strands become replication
fork.
3. Elongation of new DNA by
DNA polymerase
4. Termination of replication
and Linking of new
nucleotides to each leading
and lagging strand.
 1. Initiation - Preparing the DNA template
+Helicase unwinds DNA forming a “replication fork”at the starting point.
+Splitting happens occur at location of the chain which are rich in A-T (2 hydrogen bonds).
+Multiple replication forks along a DNA molecule create replication bubbles.
+The sites where this process first occurs are called replication origins.
2. Unwinding Process
 Helix destabilizing protein
Replication fork /SSB

DNA helicase
Topoisomerase

These two strands serve as the template for the leading and lagging strands, which will be created as DNA
polymerase matches complementary nucleotides to the templates, replication occurs in two directions.
3. Elongation Process – DNA Synthesized
/ RNA Primase adds a complimentary RNA primer to each template strand as a starting point for
replication
/ DNA Polymerase III reads the template strand (3’ to 5’) and adds new complimentary nucleotides (5’ to
3’).
/ DNA synthesized in the direction of the replication fork is called the leading strand.
Leading strands
 3’
3’

Leading strand
5’

3’

Lagging strand

5’

5’
 DNA
Polymerase I

DNA Ligase
A Summary of DNA Replication
Leading vs. Lagging
Leading Strand Lagging Strand
DNA Polymerase lll lll, l
Reading Direction 3’ – 5’ 3’ – 5’
DNA synthesis 5’ – 3’ (continuous) 5’ – 3’ (discontinuous)
direction
DNA Primase Yes (1 only) Yes (many)
RNA Primer 1 1 for each Okizaki
Fragment
Okazaki Fragments No Yes
DNA Ligase No Yes (joining Okizaki
Fragment
 4. Termination Process
/ The last step of DNA Replication.
/ RNA primers are removed by DNA polymerase I.
/ Occur when the DNA Polymerase I reaches to an end of the strands (lagging strand).
/ When the RNA primer is removed no new DNA nucleotide added because last primer
binds isn't replicated, DNA Polymerase cannot seal any gap.
/ The DNA Replication is not completed before a mechanism of repair fixes possible
errors caused during the replication.
/ Enzymes like nucleases remove the wrong nucleotides and the DNA Polymerase fills
the gaps.
Each new DNA molecule is rewound by helicase and they are
identical.
Overview the roles of transcription and translation in the flow of genetic
information
Explain transcription
Describe the stages involved:
i. initiation
ii. elongation
iii. termination
State the formation of mRNA strand from 5’ to 3’
Describe the relationship between base sequences in codons with specific amino
acids using genetic code table.
DNA Replication
Overview the roles of
transcription and translation
DNA in the flow of genetic
information

Transcription

mRNA Protein
Translation
Protein Synthesis
 Gene Expression
 DNA mRNA proteins

transcription translation

 DNA: approx only 20% is used to code for proteins

the remainder (80%) maybe inactive or
redundant or serve as initiation, termination
sites, introns, telomeres etc
Importance of Protein Synthesis
Including functional and structural
proteins.
+ Structural proteins are used for structural purposes
and dominated by the secondary protein structure
and are often fibrous.
+ Example: Collagen in connective tissues, actin and
myosin in muscles.
+ Functional proteins are used to "make things
happen” and dominated by the tertiary structure
and are often globulars.
+ Example: Enzymes are functional proteins that reduces
the activation energy in order to speed up reaction;
hormone involved in cell signalling and homeostasis,
antibodies involved in body defence mechanism.
TRANSCRIPTION
THE INFORMATION FOR PROTEIN
SYNTHESIS IS IN THE DNA IN THE
NUCLEUS.

THE INFO FROM THE DNA IS

COPIED INTO m RNA, WHICH LEAVE
THE NUCLEUS TO THE RIBOSOMES
IN THE CYTOPLASM.

THE PROTEINS ARE MADE IN THE

CYTOPLASM IN THE RIBOSOMES
Transcription
ø Transcription = the synthesis of mRNA from a DNA
template.
ø Occurs in the 5’→3’ direction. (if you don’t know what
this means go back and look it up!!)
ø Involves enzyme RNA polymerase.
ø mRNA, tRNA and rRNA must all be transcribed for protein
synthesis to take place.
ø mRNA
ø the sequence of mRNA nucleotides determine the primary
sequence of the polypeptides.
ø tRNA
ø carries the amino acids to mRNA and folds in on itself.
ø rRNA
ø major components of ribosomes.
ø Transcription occurs in 3 distinct stages:
ø Always start with Initiation
ø Proceeds with Elongation
ø Must end with Termination

ø The Chef:
ø RNA polymerase
ø It can initiate new RNA chains without a primer
ø Act as helicase, polymerase, exonuclease
Transcription (Initiation)
➢ RNA polymerase binds to the promoter site.
➢ Determines which strand of DNA will serve as the
template.
➢ Promoter = region of DNA where RNA polymerase
attaches and initiates transcription.
➢ TATA box :
➢ promoter DNA sequence.
➢ RNA polymerase binding site.

➢ the actual sequence is 5'-TATAAA-3‘ .

➢Transcription initiation complex -the area where
transcription factors and RNA polymerase are bound
to the promoter.
➢ RNA polymerase -hooks together RNA nucleotides
as they base pair along the DNA template.
➢ Transcription unit -area of DNA that will be
transcribed.
➢After polymerase is bound to the promoter DNA, the
2 DNA strands unwind and the enzyme starts
transcribing at the template strand.
 TRANSCRIPTION: The Process…..
RNA polymerase initiates RNA activated
transcription by binding to nucleotides pair
promoter at the 3' end of with the
DNA, unwinds and unzips it. complementary
bases of the
DNA strand

RNA polymerase, binds

the RNA nucleotides
together to form the
mRNA poynucleotide.
Only 1 strand of
DNA in a gene gets
transcribed.
 Initiation and Elongation

 RNA polymerase recognize and attaches to promoter site on DNA.

 Enzyme begins to separate the DNA strand .
 Segment of DNA strand unwind.
 RNA polymerase moves along DNA template.
 It unwinds 10-20 DNA bases at a time.
 RNA polymerase adds nucleotides in the 5’→3’ direction.
 As RNA polymerase moves along, the DNA double helix reforms.
 The new section of RNA ‘peels away’ as the double helix reforms
 Elongation

Encoding gene adding new RNA nucleotides in the 5’ to 3’ direction and

complimentary to the DNA template.
Works at up to 60 nucleotides/second.
 Termination

 Transcription stops when RNA polymerase reaches a section of DNA called the terminator site.
 Terminator sequence = AAUAAA
 The product is called immature or pre-mRNA
 Terminator site causes the RNA polymerase to stop transcribing DNA and release the mRNA.
 RNA polymerase detaches from the DNA
 mRNA will leave the nucleus through the nucleus pore to the cytoplasm.
 The RNA strand will go through more processing.
Sense vs. Antisense DNA strands
➢ The DNA double helix has two strands
➢ Only one of them is transcribed
➢ The transcribed strand is the antisense strand
➢ The non transcribed strand is the sense strand
➢ mRNA is complementary to the antisense strand
➢ The 5’ end of the RNA nucleotides are added to the 3’
end of the growing chain.
➢ RNA nucleotides are linked together in the same
fashion as DNA molecules.
 RNA splicing (in eukaryotes)
û In eukaryotes RNA transcription the entire gene is copied into a pre-mRNA which
never leaves the cell’s nucleus.
û Each end of a pre-mRNA molecule is modified in a particular way:
û 5’ end receives a modified nucleotide 5’ cap
û 3’ end gets a poly-A tail

û Most eukaryotic genes have interrupted coding sequence, exons and introns.
û Exon : coding regions of nucleic acids that codes for parts or all of the gene product
and is therefore expressed in mature mRNA.
û Intron : non-coding sections of nucleic acid found between coding regions that
does not code for gene product.
û It usually transcribed in eukaryotes into pre-mRNA but subsequently removed
from transcript before translation.
û During the process of RNA splicing, introns are removed and exons joined to form
a continuous protein-coding message.
û RNA splicing is carried out by spliceosomes.

Intron Exon Intron Exon Intron

Pre-mRNA mRNA
 Prokaryotes vs. Eukaryotes
 Prokaryotes
 A gene that codes for particular protein in prokaryotes produces
mRNA that begins to direct the protein synthesis as soon as it is
transcribed.

 Eukaryotes
 Pre-mRNA will be produced first (pre-mRNA wont be able to
direct protein sysnthesis).
 Pre-mRNA will then modified becoming mRNA (able to direct
protein sysnthesis).

 Why??
 Because prokaryotes DNAs are within its cytoplasm while
eukaryotes DNAs are separated from the cytoplasm by the nuclear
envelope.
Gene Information Processing

Prokaryotes Eukaryotes
 In prokaryotes, mRNA can be translated right away.
 In eukaryotes, things are not that straight forward.
 The primary transcript produced will be in form of pre-
mRNA.
 Pre-mRNA is still not a usable messenger cause it is
‘tagged’ at both ends – in form of poly-A-tail.
 Most pre-mRNA is 6000 bases long, only 1/3 is actually
transcribed, the rest are excess bases.
 Region of these non-coding RNA is called introns,
region that codes for protein is called exons.
Genetic Code
◼ Genetic code: Base triplet in DNA provides a template for ordering the
complementary triplet in mRNA molecule.
◼ Every base triplet is code for ONE amino acid.

◼ Three bases of an mRNA codon are designated as first, second and third bases.

Genetic code

1st 2nd 3rd

A T G G C A T G G C

DNA
❖ There are only FOUR nucleotide bases, to specify 20 amino acids;

❖ A-adenine, C-cytosine, G-guanine, T-thymine (unique to DNA), U-

uracil (unique to RNA) [pyrimidine, very similar to thymine].

❖ Flow of information from gene to protein is based on triplet code.

GENETIC CODE

Even though there are only 20 amino

acids that exist, there are actually 64
possible tRNA molecules:
4 X 4 X 4 = 64 possible combinations

1º NUCLEOTIDE
3º NUCLEOTIDE

CODON
CODON CODON
G C U = Gly

2º NUCLEOTIDE
❖ Consist of triplet bases (3 bases).
❖ Almost universal.
❖ Non-overlapping
❖ Commaless
❖ One codon code for one amino acid
❖ Codon are complementary to anticodon.
❖ Degenerate / one amino acids can be coded by several
codons.
❖ Start codon AUG.
❖ Stop codon UAG, UAA, UGA.
❖ Customarily written in 5’ to 3’ direction
Initiation codon
❑ Codon AUG is a starter to the process of translation.
❑ Codon AUG has dual function, as a start signal / initiation codon and it
also code for amino acid methionine (Met).
❑ Polypeptide chains begin with methionine
❑ An enzyme may subsequently remove starter amino acid from chain.
Termination codon

◼ Three triplet bases of STOP signal: UAA, UAG, UGA.

◼ marking the end of a genetic code , and the completed polypeptide chain is
released from the ribosome..

◼ Genetic massages begin with the mRNA codon AUG, which signals the
protein- synthesizing machinery to begin translating the mRNA at the
location.
Translation
 The actual process of protein synthesis.
 Translating genetic message forming of a polypeptide
uses mRNA as a template for amino acid sequence.
 mRNA translated from 5’ – 3’ direction
 Begins after mRNA enters cytoplasm.
 Uses tRNA (the interpreter of mRNA).
 Polypeptide made in the N-terminal-to-C-terminal
direction.
 3 steps (initiation, elongation and termination).
Structures that involved in translation

Facilitate the
specific coupling of
tRNA anticodons
with mRNA codons
during protein
synthesis.

tRNA transfer amino

acid from cytoplasm to
ribosome

Carries genetic information that

attach to ribosomes which translates
codon into amino acids.
 Messenger RNA (mRNA)

+ Single strand RNA nucleotides.

+ Synthesized in transcription.
+ Composed of codons.
+ Codons are 3-base sequences of mRNA.
Ribosomal RNA (rRNA)
 Made of proteins and rRNA.
 Each has a large and small subunit.
 60s and 40s in eukaryotes, 1 rRNA + 33 ribosomal proteins
(RP) in small and 4 rRNA + 49 RP in large subunits
 50s and 30s in prokaryotes, 1 rRNA + 21 RP in small and 3
rRNA + 31 RP in large subunits
 Each has three binding sites for tRNA on its surface.
 Each has one binding site for mRNA.
 Facilitates codon and anticodon bonding.
 Components of ribosomes are made in the nucleus and
exported to the cytoplasm where they join to form one
functional unit.
 The three tRNA binding sites are:
1. A site=holds tRNA that is carrying the next amino
acid to be added
2. P site= holds tRNA that is carrying the growing
polypeptide chain
3. E site= where discharged tRNAs leave the ribosome
tRNA
 tRNA is transcribed in the nucleus and must enter the cytoplasm.

 Carries amino acids to the ribosome.

 During tRNA charging each tRNA picks up an amino acid from the cytoplasm.

 tRNA molecules are used repeatedly.

 Each tRNA molecule links to a particular mRNA codon with a particular amino acid.

 When tRNA arrives at the ribosome it has a specific amino acid on one end and an
anticodon on the other.
 Anticodons (tRNA) bond to codons (mRNA).
tRNA Structure
tRNA Activation
Via process called Aminoacylation (charging).
Done by aminoacyl-tRNA-synthetase (AtS).
Produces aminoacyl-tRNA (charged-tRNA) and
using energy of ATP.
20 different AtS for 20 different amino acids.
 Each enzyme recognizes particular structural
features of the tRNA it aminocylates.
 tRNA molecule is reactivated many times
(recycled).
 Aminoacylation
+ Enzyme aminoacyl-tRNA synthetase
(AtS):
+ Attach appropriate amino acid to tRNA.

+ Hydrolyses ATP to AMP which join

amino acid to form aminoacyl-AMP.
+ Uncharged tRNA molecule binds to AtS.

+ AtS transfers the aminoacyl-AMP to the

tRNA displaces the AMP.
+ tRNA + amino acid = aminoacyl-tRNA

+ AtS then releases the charged-tRNA.

+ Covalent linkage between amino

acid and adenine at 3’ end of tRNA.

Translation
TRANSLATION: INITIATION
 Small ribosomal subunit recognizes and binds to
mRNA at 5’ cap.
 Small subunit moves along 5’ leader of mRNA until
it reach the start codon (AUG).
 Once ribosome small subunit reach AUG, initiator tRNA(charged
tRNA) attaches AUG.
 Hydrogen bond forms between start codon AUG and anticodon with
Methionine at a P site.
 Then the larger subunit comes over the tRNA so that it is in the P
(middle) position in the ribosome.
 The A site will remain vacant and ready for the other aminoacyl-tRNA.
TRANSLATION: ELONGATION
 A peptide bond is formed between the new amino acid in
the A site and the growing polypeptide in the P site.
➢ Codon recognition.
➢Peptide bond formation catalyzed by Peptidyl transferase located in the large subunit.
➢Translocation: ribosome moves along mRNA in the 5’→3’ direction, aminoacyl tRNA shifts from A site to P
site.
➢The old tRNA has just gone through the cycle EBME (enter, bond, move, exit).
➢This process continues until the there is a stop codon.
 TRANSLATION: TERMINATION
 Triggered when a stop codon (or nonsense codons) UAA, UAG,
UGA enters the A site.
 These codons
 Do not code for any amino acid.
 Do not bind with any tRNA.
 Bind with a cytoplasmic protein called the releasing factor (RF)
 Release factor:
 Adds water molecule instead of amino acid to polypeptide.
 Hydrolyses bond between the polypeptide chain and the tRNA in
the P site.
 Polypeptide is released.
 Translation complex disassembles.
Polyribosome or Polysome
1. Explain the concept of operon and gene
regulation
2. Describe the mechanism of the lac operon in
the absence and presence of lactose.
Gene regulation
 Is the process of turning genes on and off.
 During early development, cells begin to take on specific
functions.
 Ensures that the appropriate genes are expressed at the
proper times.
 Can also help an organism respond to its environment.
 In other words gene regulation is the modulation any of
the stages of gene expression.
Concept of operon
 They are only found in prokaryotes.
 An operon is a group of genes that are transcribed
at the same time.
 They usually control an important biochemical
process.
 Coordinated regulation unit of transcription in
bacteria.
 The system consists of a regulator gene and an
operon.
 Transcription of an operon produces mRNAs for 3
polypeptides which produced enzymes in lac operon.
Lac operon consists of:

OPERON
 The lac operon consists of:
 Promoter
 RNA polymerase binding site + initiation site

 Operator
 Repressor protein binding site.
 Functions as a 'switch’ which activates or deactivates a
sequence of structural genes.
 Structural genes with related functions
 Gene which has the genetic information for determine
sequence of amino acid.
 Which enzyme to digest lactose.

 Regulator/Repressor genes
 Codes for and results in production of repressor protein
that bind to the operator.
 Protein synthesis involved 2 stages…?
 Lac operon - gene regulation and
expression happen at the transcription
stage.
 Translation for lac operon will produce
polypeptides – enzymes – hydrolyse lactose.
The lac Operon
• Is the section of DNA that acts as on/off switch for genes that
control the metabolisme of lactose.
• When lactose is present, lac operon is switched on and
produces enzymes.
 Found in E.coli. consists of a promoter, an operator regulatory
gene and 3 structural genes.
 Regulatory gene (lac I gene) encodes for the synthesis of an
active repressor.
Adapting to the environment
 E. coli can use either glucose, which is a
monosaccharide, or lactose, which is a disaccharide.
 However, lactose needs to be hydrolysed (digested)
first.
 The cell will produced a new enzyme - -galactosidase.
 Only produced when it was needed.
 So the bacterium prefers to use glucose when it can.
1. When lactose is absent
 A repressor protein is active and binds to the operator makes
the operon deactivate.
 This blocks the binding of RNA polymerase to the promoter,
preventing transcription.

Repressor RNA
protein Blocked polymerase

DNA
I
O z y a
Regulator Operator
lac operon
gene site
© 2007 Paul Billiet ODWS
 2. When lactose is present
 An isomer of lactose allolactose is formed and acts as inducer
and binds to the another active site of repressor protein
(allosteric site).
 This causes the repressor protein to alter its shape (a
conformational change) and becomes inactive. It can no
longer sit on the operator site.
 RNA polymerase can now reach its promoter site.
 Transcription can occur and -galactosidase, permease and
transacetylase enzymes are produced.

DNA

I O z y a
 2. When lactose is present
 An isomer of lactose allolactose is formed and acts as inducer and
binds to the another active site of repressor protein (allosteric site).
 This causes the repressor protein to alter its shape (a
conformational change) and becomes inactive. It can no longer sit
on the operator site.
 RNA polymerase can now reach its promoter site.
 Transcription can occur and -galactosidase, permease and
transacetylase enzymes are produced.

DNA
I O z y a
Promotor site
© 2007 Paul Billiet ODWS
 RNA polymerase binds to the promoter and 3 structural
genes are transcribed.
 The mRNA is translated into enzymes that aid in lactose
digestion.

LacZ β-galactosidase Hydrolyses lactose to glucose

and galactose.
LacY Permease Encodes a cell membrane
(lactose protein to transport Lactose
permease) across the cell wall.
LacA Transacetylase Transfer an acetly group from
acetyl CoA to β-galactosidase.