2nd Group:

Farih Abdillah
Petir Langlang Buana
Feri Eko Hermanto
Wahyu Isnia Adharini
Sayyidatul Awalia Nuzula
Meylinda Kurniawati

(135090107111010)
(1350901xxxxxxxx)
(145090100111001)
(145090100111002)
(145090100111008)
(145090100111004)

Introduction

Replication Model Theories

Conservative Model
Parental molecule directs synthesis of an entirely new
double-stranded molecule, such that after one round
of replication, one molecule is conserved as two old
strands. This is repeated in the second round.

Fig. 2. Conservative model of DNA replication (Carr, 2011)

Replication Model Theories

Semi-conservative Model
Two parental strands separate and each makes a
copy of itself. After one round of replication, the two
daughter molecules each comprises one old and one
new strand. Note that after two rounds, two of
the DNA molecules consist only of new material,
while the other two contain one old and one new
strand.

Fig. 3. Semi-Conservative model of DNA replication (Carr, 2011)

Replication Model Theories

Dispersive Model
Material in the two parental strands is distributed
more or less randomly between two daughter
molecules. In the model shown here, old material is
distributed between the two daughters molecules.

Fig. 4. Dispersive model of DNA replication (Carr, 2011)

Replication Components
1.
2.
3.
4.
5.

DNA
Helicase
Topoisomerase and Gyrase
Single stranded binding protein (SSB)
DNA polymerase
Eukaryotic
: , , , ,
Prokaryotic : I, II and III
6. Primase
7. Ligase

Replication Components

Replication Mechanism
Initiation
Begin at specific site called origin of replication (Ori)
to which certain initiator proteins bind and trigger
unwinding
Enzymes known as Helicases unwind the double
helix by breaking the hydrogen bonds between
complementary base pairs
Other proteins called SSB keep the single strands
from rejoining and finally form a replication fork
The topoisomerase proteins surround the unzipping
strands and relax the twisting that might damage the
unwinding DNA.
Fig. 4. Initiation process of DNA replication (Reece et al., 2014)

Replication Mechanism
Elongation
Specialized RNA polymerase called primase initiates
polynucleotide synthesis and by creating a short RNA
polynucleotide strand complementary to template DNA
strand
DNA polymerase extends the new strand with
nucleotides complementary to the template DNA
DNA gyrase also is required so untwists the rotations that
are generated ahead of the moving replication fork
RNA nucleotides in the primer are removed and replaced
with DNA nucleotides by DNA polymerase I
The gaps are closed with the action DNA Ligase (adds
phosphate in the remaining gaps of the phosphate sugar backbone)
Fig. 4. Elongation stage of DNA replication (Campbell et al., 2010)

Replication Mechanism
Leading and Lagging Strand
Leading Strand
The new DNA strand at the replication fork is
synthesized in the 5 ' 3' continuously
Lagging Strand
DNA strands on the opposite side of the Leading
Strand running in the 3 'to 5', because the DNA
polymerase III is unable to synthesize in the 3 ' 5'
Lagging Strand being synthesized in short segments
known as Okazaki fragments

Replication Mechanism
Leading and Lagging Strand

Replication Mechanism
Termination
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
helped by Ligase

Fig. 4. Mismatch repairing by nuclease (Reece et al., 2014)

Replication Mechanism
Termination
Linear DNA cant complete the 5 ends of daughter
DNA strands, especially in lagging strand
As a result, repeated rounds of replication produce
shorter and shorter DNA molecules with uneven
ends
So Eukaryotic chromosomal DNA molecules have
special nucleotide sequences called telomeres at
their ends and An enzyme called telomerase
catalyzes the lengthening of telomeres
Telomerase is not active in most human somatic
cells, but its activity varies from tissue to tissue
Fig. 4. Shortening of the ends of linear DNA molecules (Reece et al., 2014)

Differences of DNA Replication between

Procaryotic and Eucaryotic Cells
Prokaryotic
Place

Eukaryotic

Cytoplasm

Nucleus

DNA Polymerase

DNA polymerase I and II: DNA repair

DNA polymerase III: DNA synthesis

DNA polymerase : Priming of replication of both strands

DNA polymerase : Elongation of lagging strand
DNA polymerase : DNA repair
DNA polymerase : Elongation of leading strand
DNA polymerase : Replication of mitochondrial DNA

Primer synthesis

requires a primosome composed of the DNA

carried out by a primase associated with the host DNA
helicase, DnaB, and the primase, DnaG
polymerase
(in E. coli)

Speed of Replication and fork

replication movement

Faster

Slower

The origin site of replication

Only one origin per DNA molecule

Multiple origin in each eaukaryotic chromosome

Strand synthesis

Leading and Lagging strand synthesis by DNA

polymerase III

Leading strand is synthesized by DNA polymerase and

lagging strand is synthesized by DNA polymerase which
is made in short pieces

Termination

No telomerase, spesific termination siter or

ter sites are present

Termination of replication in linear chromosomes involves

synthesis of telomer and the enzyme involved is
telomerase

Polymerase Chain Reaction (PCR)

PCR (Polymerase Chain Reaction) is an enzymatic
method for the amplification of DNA by in vitro.
PCR was first developed in 1985 by Kary B. Mullis.

Polymerase Chain Reaction (PCR)

Components
DNA template
Oligonucleotide primer
Deoxyribonucleotide triphospate (dNTP)
Enzyme DNA Polymerase
Buffer Compound

Polymerase Chain Reaction (PCR)

Principle and Methods
Denaturation
: 2 hour, 40 minute, 5 minute
each at a temperature of 92,5; 95 and 97,5 C.
Annealing
: 50 - 60 C for 30 45 second.
Extension
: 72 C to produce 32 100
nucleotides/sec

PCR cycle is repeated 25-30 times

PCR Mechanism

Polymerase Chain Reaction (PCR)

Types of PCR
1. Restriction Fragment Length Polymorphism (RFLP)
2. Inverse-PCR
3. Nested-PCR
4. Quantitative-PCR
5. Reverse Transcriptase (RT-PCR)
6. Random Amplified Polymorphic DNA (RAPD)

Conclusion
1. DNA replication model theories are Conservative, Semiconservative, and
Dispersive. The most acceptable model based on replication mechanism is
Semiconservative model
2. DNA replication mechanism have 3 major stages, they are Initiation,
Elongation and Termination
3. The Mechanism of DNA replication between Prokaryotic and Eukaryotic
cells doesnt have any significant differences, only based on their enzymes
4. Mechanism of PCR is adapted from DNA replication. There are 3 steps of
PCR methods, denaturation, annealing and extension.