DNA REPLICATION AND PROTEIN

SYNTHESIS, GENETIC
ENGINEERING , RECOMBINANT
DNA
DNA Replication 

In the process of DNA replication, the DNA makes

multiple copies of itself. It is a biological polymerisation,
which proceeds in the sequence of initiation, elongation,
and termination. It is an enzyme-catalysed reaction. DNA
Polymerase is the main enzyme in the replication
process.
PROPERTIES OF DNA REPLICATION:

1. DNA REPLICATION IS SEMICONSERVATIVE

2. THE TWO DNA STRANDS ARE ANTI PARALLEL

3. DNA HAS COMPLEMENTARY BASE PAIRING

4. H- BONDS HOLDS NUCLEOTIDES TOGETHER

 1. DNA REPLICATION USUALLY PROCEEDS BIDIRECTIONALLY.
STEPS IN DNA 2. HELICASE SEPARTAES THE DNA STRANDS.
REPLICATION: 3. TOPOISOMERS REDUCES TORSIONAL STRAIN & POSITIVE
SUPERCOILS.
4. SSB PROTEINS STABILIZES THE ISOLATED STRANDS.
5. DNA REPLICATION REQUIRES A RNA PRIMER TO BEGIN.
6. DNA POLYMERASE III ADDS NUCLEOTIDES IN THE 5’ TO 3’
DIRECTION.
7. DNA REPLICATION IS SEMI DISCONTINUOUS.
8. DNA PLOYMERASE 1 REMOVES RNA PRIMER & REPLACES IT
WITH DNA.
9. DNA LIGASE SEALS THE NICK AFTER THE PRIMER IS
REPLACED WITH DNA.
10.DNA POL. 1 & 111 HAVE 3’ TO 5’ EXONUCLEASE ACTIVITY
(PROOFREADING).
11.DNA POL. 1 HAVE 5’ TO 3’ EXONUCLEASE ACTIVITY ( DNA
REPAIR).
DNA Replication Steps
Following are the important steps involved in DNA replication:

1. Initiation
DNA replication demands a high degree of accuracy because even a minute
mistake would result in mutations. Thus, replication cannot initiate randomly at
any point in DNA.

For the replication to begin there is a particular region called the

origin of replication-This is the point where the replication originates. Replication
begins with the spotting of this origin followed by the unwinding of the two DNA
strands.
Unzipping of DNA strands in their entire length is not feasible due to high energy
input. Hence, first, a replication fork is created catalysed by the helicase
enzyme, which unzips the DNA strand.
2. Elongation
As the strands are separated, the polymerase enzymes start synthesising
the complementary sequence in each of the strands. The parental strands
will act as a template for newly synthesising daughter strands.
It is to be noted that elongation is unidirectional i.e. DNA is always
polymerised only in the 5′ to 3′ direction. 
Therefore, in one strand (the template 3‘→5‘) it is continuous, hence called
continuous replication/leading strand.
while on the other strand (the template 5‘→3‘) it is discontinuous
replication/lagging strand. They occur as fragments called Okazaki
fragments. The enzyme called DNA ligase joins them later.
3.Termination
Termination of replication occurs in different ways in different organisms.

In E.coli like organisms, chromosomes are circular. And this happens when

the two replication forks between the two terminals meet each other.
Role of Enzymes in DNA Replication
DNA replication is a highly enzyme-dependent process. There are many
enzymes involved in DNA replication, which includes the enzymes,

• DNA-dependent DNA polymerase,

• helicase,
• ligase, etc. 
• Among them, DNA-dependent DNA polymerase is the main enzyme.

DNA-dependent DNA polymerase (enzyme)

It helps in the polymerisation, catalyses and regularises the whole process
of DNA replication with the support of other enzymes..
Helicase
Helicase is the enzyme, which unzips the DNA strands by
breaking the hydrogen bonds between them. Thus, it helps
in the formation of the replication fork.
2. Elongation( review)
As the strands are separated, the polymerase enzymes start synthesising
the complementary sequence in each of the strands. The parental strands
will act as a template for newly synthesising daughter strands.
It is to be noted that elongation is unidirectional i.e. DNA is always
polymerised only in the 5′ to 3′ direction. 
Therefore, in one strand (the template 3‘→5‘) it is continuous, hence called
continuous replication/ leading strand .
while on the other strand (the template 5‘→3‘) it is discontinuous
replication/lagging strand. They occur as fragments called Okazaki
fragments. The enzyme called DNA ligase joins them later.
Ligase (enzyme)
Ligase is the enzyme which joins together the Okazaki fragments of the
discontinuous DNA strands.
Primase (enzyme)
This enzyme helps in the synthesis of RNA primer complementary to the
DNA template strand.
Single-stranded Binding Proteins(enzymes)
It binds to single-stranded DNA and protects it from forming secondary
structures.
Protein Synthesis – Central Dogma
The central dogma describes how the DNA makes its own copies by DNA
replication, then codes for the RNA by means of a transcription and RNA
codes for the proteins by translation.

For the creation of a new generation, the genetic information stored in

the DNA has to be transferred from one generation to another. The
process begins with the replication of DNA followed by transcription and
translation. Let us have a discussion on translation eg.protein synthesis
in cell and the structure of tRNA.
Structure of tRNA

The DNA is transcribed into mRNA on the basis of their

complementarity. Translation is the process of conversion of
nucleic acid information into amino acids. There is no
complementarity between amino acids and mRNA. Hence,
translation is not controlled by complementarity but by the 
genetic code. Since amino acids cannot read this genetic code, they
are dependent on an adapter molecule. This adapter molecule is
called tRNA (transfer RNA).
Transcription is the process by which the information in a
strand of DNA is copied into a new molecule called messenger RNA
(mRNA). 
∙ The segments of DNA are transcribed into RNA molecules that
encode proteins are said to produce messenger RNA.

∙ This mRNA encodes genes that are translated into protein by

the process of translation. 

∙  Other segments of DNA are copied into RNA molecules called non-coding
RNAs.
TRANSCRIPTION-
is the process by which the information in a
strand of DNA is copied into a new molecule
called messenger RNA (mRNA). 
GENETIC ENGINEERING

Genetic engineering, also called genetic modification, is the direct

manipulation of an organism’s genome using biotechnology. It is a
set of technologies used to change the genetic makeup of cells,
including the transfer of genes within and across species boundaries
to produce improved or novel organisms.
STEPS IN
GENETIC
ENGINEERING
Recombinant DNA, or rDNA,
is DNA that is formed by combining DNA from
different sources through a process called genetic
recombination. Often, the sources are from different
organisms. Generally speaking, DNA from different
organisms has the same general chemical structure.
Recombinant DNA technology- genetic
material from multiple sources….. is the
process used to create a recombinant DNA molecule which carries
the DNA of interest and vectorgenetic
DNA while
engineering –is the process of using
recombinant DNA to alter genEtic make up
of an organism; is a broad term used to describe the
processes involved in manipulation of the genetic structure of an
organism.  Genetic engineering is done under in
vitro conditions (outside a living organism, under a controlled
environment).
For example, one of the most common GMO crops is Bt corn,
which is genetically modified to produce the insecticide Bt
toxin. By making this toxin, the corn is able to resist pests,
reducing the need for pesticides (3Trusted Source).

GMO crops are incredibly common in the United States, with

at least 90% of soy, cotton, and corn being grown through
genetic techniques (4Trusted Source).
In fact, it’s estimated that up to 80% of foods in supermarkets contain
ingredients that come from genetically modified crops.

While GMO crops make farming much easier, there is some concern around
their potential effect on the environment and their safety for human
consumption — specifically surrounding illnesses and allergies (
5Trusted Source).

However, the Food and Drug Administration (FDA), Environmental

Protection Agency (EPA), and USDA maintain that GMOs are safe for
human and animal consumption (6Trusted Source).
In fact, an analysis of 147 studies from 2014 found that GMO technology has reduced chemical
pesticide use by 37% and increased crop yields by 22% (8Trusted Source).

Other GMO crops have been modified with genes that help them survive stressful conditions,
such as droughts, and resist diseases like blights, resulting in a higher yield for farmers (
9Trusted Source, 10Trusted Source, 11Trusted Source).

Together, these factors help lower the costs for the farmers and consumers because it allows a
greater crop yield and growth through harsher conditions.

Additionally, genetic modification can increase the nutritional value of foods. For example, rice
high in beta carotene, also called golden rice, was developed to help prevent blindness in
regions where local diets are chronically deficient in vitamin A (12Trusted Source).

Moreover, genetic modification may be used simply to enhance the flavor and appearance of
foods, such as the non-browning apple (13Trusted Source).

In addition, current research suggests that GMO foods are safe for consumption (
14Trusted Source).