DNA replication; Applications / Developments in Genetic Engineering

DNA replication makes the transfer of genetic information from one generation to another possible
It is an important phenomenon happening inside our cells, that allows the body to maintain homeostasis and integrity
of the body

With the available information about DNA replication, scientific communities today have a proper idea of genome
sequencing which has now been applied in different areas ranging from clinical diagnosis to possible treatment of
genetic diseases

DNA replication has made sequencing of whole human genome sequencing possible
Cloning of genes has also been possible by DNA replication

Enzymes involved in DNA replication have now been greatly studied due to their wider applications. The recent
breakthrough Cas9/ CRISPR technology where nucleases are used to cleave the desired portion of DNA and replace it
with required nucleotides is the prime example of how we can use these enzymes and make potential advancements in
them thus broadening and exploring their uses
Polymerase Chain Reaction uses DNA polymerases to repeatedly replicate DNA in-vitro and has numerous
applications in diagnosis, sequencing, and recombinant DNA technology

The formation of complementary DNA (cDNA) can also be considered as an example of a wider application of the
enzymes involved in DNA replication

There are various applications of DNA replication, we can even consider that if there is any technique involving genes,
some way or the other DNA replication is applied
Similarities between Prokaryotic and Eukaryotic DNA Replication

- The unwinding mechanism of DNA before replication is initiated is the same for both Prokaryotes and eukaryotes

- In both organisms, the DNA polymerase enzyme coordinated the synthesis of new DNA strands

- Additionally, both organisms use the semi-conservative replication pattern, making the leading and lagging strands in
different directions. Okasaki fragments make the lagging strand

- Lastly, both organisms initiate DNA replication using a short RNA primer
S.N Pol a, e, d/ b, g
Eukaryotic DNA Replication (in eukaryotic cells) Prokaryotic DNA replication (in prokaryotic cells)
.
This process takes place in the cell’s nucleus.
1. This process takes place in the cell’s cytoplasm.

With multiple sites for the origin of replication per DNA molecule
2. Only a single site for the origin of replication per DNA molecule.

Initiation of DNA replication is carried out by multi-subunit Initiation of DNA replication is carried out by protein DnaA and
3.
proteins, origin recognition complex. DnaB.

4. Multiple replication forks are formed in a DNA molecule. Only two replication forks are formed in a DNA molecule.

Okazaki fragments are short of around 100-200 NTDs in length Okazaki fragments are large, around 1000-2000 NTDs in
5.
length.
6. It is a slow process with around 100 NTDs added per second. It is a fast process with around 2000 NTDs added per second.

7. DNA is linear and double-stranded. DNA is circular and double-stranded.

DNA polymerase involved in eukaryotic DNA replication is DNA DNA polymerase involved in prokaryotic DNA replication is DNA
8.
polymerases ε, α, and δ polymerase Ι, and ΙΙΙ
Euk-cell has telomeres at the end of DNA thus they are
9. Prokaryotic cells have circular DNA thus they are not replicated.
replicated.

10. DNA gyrase (Telomerase) is needed. DNA gyrase (Telomerase) is not needed.
 In the semi-
conservative model,
the 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 In the conservative model, the parental molecule directs synthesis of an entirely
strand. 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.
In the dispersive model, material in the two parental strands is distributed more or
2010 PJ Russell, iGenetics 3rd ed.; all text
material © 2011 by Steven M. Carr
less randomly between two daughter molecules. In the model shown here, old
https://www.mun.ca/biology/scarr/iGen3_03- material is distributed symmetrically between the two daughters molecules. Other
01.html distributions are possible.
 Semiconservative model of Replication
- Each double helix chain/ strand serving as the template for synthesizing the other
- For the replication of DNA molecules, Watson and Crick proposed that replication involved the disruption of hydrogen
bonds followed by a rotation and separation of the two polynucleotide strands.
-Each purine and pyrimidine base of each polynucleotide strand attracts a complementary free nucleotide
available for polymerization in the cell, and to hold it in place means of the specific hydrogen bonds

- Once held in place on the parent template chain, the free nucleotides are linked together by forming the
phosphodiester bonds that linked adjacent deoxyribose residues, forming a new polynucleotide molecule of a
predetermined base sequence.

- The result of replication is two double-stranded DNA molecules with sequences identical to the original one.
- The original left strand is present in one of these daughter molecules, while the original right strand is in the
other
- Since each progeny retains half of the parent DNA molecule, this replication pattern is considered semi-
conservative
- The DNA replication in prokaryotes and eukaryotes is similar but not the same. The process is more complicated in eukaryotes than
Other models of DNA replication are equally possible. They are:

1.Conservative replication:
The new DNA molecule would consist of two freshly synthesized strands, and both of the parent double helix’s
strands are conserved

2.Dispersive replication:
The parent double helix is broken up during replication, and fragments of the parent strands are mixed with
newly synthesized strands to form the two new double helices.