:

Expression of Biological
Information
 6.0 Expression of Biological
Information

6.1 DNA and genetic information

6.2 DNA replication
6.3 Protein synthesis: transcription and
translation
6.4 Gene regulation and expression – Lac
operon
 6.0 Expression of Biological
Information

6.1 DNA and genetic information

6.2 DNA replication
6.3 Protein synthesis: transcription and
translation
6.4 Gene regulation and expression – Lac
operon
Learning Outcomes :
6.2 Describe semi-conservative replication of DNA

Models of DNA
replication

• Conservative

• Semi-conservative

• Dispersive
Learning Outcomes :
6.2 Describe semi-conservative replication of DNA

Conservative

• The parental
double helix
remains intact and
an all new copy is
made
Learning Outcomes :
6.2 Describe semi-conservative replication of DNA

Semi-conservative
• The two strands of the
parental molecule
separate, and each
function as a template
for synthesis of a new
complementary strand
Learning Outcomes :
6.2 Describe semi-conservative replication of DNA

Dispersive
• Each strand of
both daughter
molecule
contains a
mixture of old
and newly
synthesized
parts
Learning Outcomes :
6.2 Describe semi-conservative replication of DNA

Evidence for semi-conservative

hypothesis

• Experiment performed by Meselson & Stahl in

1950, using the radioisotope labelling
technique

• 2 different isotopes of Nitrogen are used :

a) 14N is the light isotope of nitrogen
b) 15N is the heavy isotope of nitrogen
Fig. 14.12b
Learning Outcomes :
6.2 Describe semi-conservative replication of DNA

Conclusion:

• The results indicates that, replication of DNA

strands by mean of semi-conservative
mechanism

• Each strand of a DNA double helix acts as a

template for the synthesis of a new strand
Learning Outcomes :
6.2 Describe DNA replication during cell division

Overview of DNA replication

Learning Outcomes :
6.2 Describe DNA replication during cell division

Origin of replication

• A special sites where the replication of

DNA molecule begins

• It is where two parental strands separate

and form replication bubbles
Learning Outcomes :
6.2 Describe DNA replication during cell division

• In bacterial chromosome
(circular DNA) has a single
origin

• In eukaryotes, DNA
replication begins at many
sites along the giant DNA
molecule of each
chromosome
Learning Outcomes :
6.2 Describe DNA replication during cell division

Replication process
The steps:

1. DNA gyrase relieves strain

of double-stranded DNA

2. A portion of the parental

double helix is unwound by a
DNA helicase.
Learning Outcomes :
6.2 Describe DNA replication during cell division

3. Molecules of single
strand binding
protein stabilize the
unwound template
strands

4. RNA primer is first

synthesized at the
origin of replication
Learning Outcomes :
6.2 Describe DNA replication during cell division

5. RNA primer is synthesized by a protein complex

known as RNA primase

6.Primers are generally 5-10 nucleotides long

Learning Outcomes :
6.2 Describe DNA replication during cell division

7. RNA primase removed, DNA polymerase

III synthesized DNA nucleotides to RNA
primer
Learning Outcomes :
6.2 Describe DNA replication during cell division

8. DNA polymerase III add nucleotides only to the free 3’

end of growing DNA strand, never to the 5’

9. Thus, a new DNA strand can elongate only in the 5’ to 3’

direction
Learning Outcomes :
6.2 Describe DNA replication during cell division

10. DNA polymerase I

then replace the
RNA primer with
DNA nucleotides

11. Finally DNA ligase

joins the 3’ end of
the DNA that
replaces the primer
to the rest of new
strand
Learning Outcomes :
6.2 Describe DNA replication during cell division

Leading and Lagging strands

• Along one template

strand, DNA polymerase
III adding nucleotides in
5’ to 3’ direction
continuously, towards
the replication fork

• The DNA strand made

by this mechanism is
called leading strand
Learning Outcomes :
6.2 Describe DNA replication during cell division

• To elongate the other

new strand of DNA, DNA
polymerase III must
work along the other
template strand in the
direction away from the
replication fork

• The DNA strand

synthesized in this
direction is called
lagging strand
Learning Outcomes :
6.2 Describe DNA replication during cell division

• In the lagging strand, only short piece

of DNA can be synthesized because
DNA polymerase III need to move far
away from the fork

• This short segment is called Okazaki

fragments

• Each Okazaki fragment initiated by

separate RNA primer
Learning Outcomes :
6.2 Describe DNA replication during cell division

 DNA polymerase III adds DNA nucleotides

to the primer forming Okazaki fragment

 After reaching the next RNA primer, DNA

polymerase III falls off

 DNA polymerase I replaces the RNA primer

with DNA nucleotide

 Finally, DNA ligase joins the fragments of

the lagging strand together