By:

Jyoti Gurung
Alisha Gurung
Diwakar Chaudhary
Nishma Bajracharya
Manish Karmacharya
Molecular genetics is the field
of biology which studies the
structure and function of genes
at a molecular level. The field
studies how the genes are
transferred from generation to
generation. Molecular genetics
employs the methods of genetics
and molecular biology.
All DNA is made up of a base consisting of
sugar, phosphate and one nitrogen base.
There are four nitrogen bases, adenine (A),
thymine (T), guanine (G), and cytosine (C).
The nitrogen bases are found in pairs, with
A & T and G & C paired together and are
complementary.
The sequence of the nitrogen bases can be
arranged in an infinite ways, and their
structure is known as the famous "double
helix“. The sugar used in DNA is
deoxyribose. The four nitrogen bases are
the same for all organisms. The sequence
and number of bases is what creates
Double helical model of
DNA
Recombinant DNA is a form of
artificial DNA that is engineered through
the combination or insertion of one or
more DNA strands, thereby combining
DNA sequences that would not normally
occur together. In terms of genetic
modification, recombinant DNA is
produced through the addition of
relevant DNA into an existing
organismal genome, such as the
plasmid of bacteria, to code for or alter
different traits for a specific purpose,
Restriction enzymes: Certain
endonucleases – the enzymes that
cut DNA at specific DNA sequences
within the molecule, are a key tool in
recombinant DNA research. These
are called restriction enzymes.
Vectors: A vector is a DNA molecule
into which foreign DNA may be
inserted which can then replicate in
an appropriate cell.
Blunt-ended DNA : Two strands of
a DNA duplex having ends that are
flush with each other.
Sticky –ended DNA:
Complementary single strands of
DNA that protrude from opposite
ends of a DNA duplex or from the
ends of different duplex molecules.
Plasmid: A small, extrachromosomal
, circular molecule of DNA that
EnzymesA Reaction Primary Use
Alkaline Dephosphorylates 5’ Removal of 5’ –PO4
phosphate ends of RNA and groups prior to kinetic
DNA ligase DNA.
Catalyzes bonds labeling
Joining oftoDNA
prevent self –
molecules.
between DNA ligation.
DNA molecules.
Synthesizes double- Synthesis of double-
polymerase I stranded DNA from stranded cDNA; nick
single stranded DNA. translation; generation of
Dnase I Under appropriate blunt ends from sticky
Nick translation; mapping
conditions, produces ends.
of hypersensitive sites;
single-stranded mapping protein-DNA
Exonuclease III nicks in DNA.
Removes interactions.
DNA sequencing;
nucleotides from 3’ mapping of
ends of DNA. hypersensitive sites;
Reverse Synthesizes DNA mapping
Synthesisprotein-DNA
of cDNA from
transcriptase from RNA template. interactions.
mRNA; RNA (5’end)
mapping studies.
 It can supply large amounts of materials that
could not be obtained by conventional purification
methods (eg., interferon, tissue plasminogen
activating factor)
 It can provide human material (insulin, growth
hormone)
 Proteins for vaccines(example: hep-B) and for
diagnostic testing.(eg. AIDS tests) can be
obtained.
 It is used for the production of recombinant
pharmaceuticals and in forensic medicine.
 It is also used to engineer plants that are more
resistant to drought or temperature extremes,
more efficient at fixing nitrogen, or that produce
seeds containing the complete complement of the
The classic example of the use of
recombinant DNA technology is sickle
cell disease which is caused by
mutation of single base out of 3x109
in the genome, a T-A DNA
substitution, which in turn results in
an A-U change in the mRNA
corresponding to the sixth codon of
the ß globin gene. The altered codons
specifies a different amino acid
i.e.valine rather than glutamic acid,
Gene therapy is being studied as a
possible treatment for sickle cell
anaemia. Researchers are looking to
see whether a normal gene can be
planted in the bone marrow of a
person with sickle cell anaemia, and
thus cause the body to produce
normal red blood cells. Researchers
also are studying the possibility of
treatment to “turn off” the sickle cell
gene or “turn on” a gene that makes
 1. Isolating Gene
The gene for producing HUMAN insulin
protein is isolated. The gene is part of the
DNA in a human chromosome. The gene
can be isolated and then copied so that
many insulin genes are available to work
with.
2. Preparing target DNA
First, a circular piece of DNA called a
plasmid is removed from a bacterial cell.
Special proteins are used to cut the
plasmid ring open.
 4. Insert ing Plasmid back into
cell
The bacterial DNA now contains the
human insulin gene and is inserted
into a bacteria. Scientists use very
small needle syringes to move the
recombined plasmid through the
bacterial cell membrane.
5. Plasmid multiply
Many plasmids with the insulin gene
are inserted into many bacterial cells.
The cells need nutrients in order to
grow, divide, and live. While they live,
6. Target Cells Reproduce
Human insulin protein molecules
produced by bacteria are gathered
and purified. The process of purifying
and producing cow and pig insulin
has been greatly reduced or
eliminated.
7. Cells Produce Proteins
Millions of people with diabetes now take
human insulin produced by bacteria or
yeast (biosynthetic insulin) that is
CLONING
Cloning is the isolation and growth of
a single, genetically uniform cell and
its offspring. Recombinant DNA
cloning is the isolation of a cell line
which contains (and replicates) a
single, unique recombinant DNA from
a pool (LIBRARY) of many different
recombinant DNA’s.
The first human clone, a healthy 7-pound
girl named Eve, was born the 26th of
December 2002, according to Clonaid, a
private company linked to the Raelian
religious sect.
A transgenic animal or Genetically
modified organism is an organism
whose genetic material has been altered
using genetic engineering techniques i.e.
DNA recombinant technology.
Scientists have produced a number of
transgenic creatures they hope will bring
major benefits to mankind. For example,
human genes have been put into pigs to
allow the pigs to produce human insulin, a
substance needed to control diabetes, one
of the fastest-growing diseases in affluent
countries.
Sheep are being altered to generate a
protein that may fight the lung
disease emphysema. Designer dairy
products are also on the drawing
boards. Geneticists hope to end up
with a breed of cow that produces, for
example, only low-fat milk.

Finally, researchers are optimistic

that they will be able to turn animals,
principally pigs, into sources of
organs for human transplants. To
Transgenic animal

Ordinary zebra fish

Gold Fish are a type of zebrafish with
recombinant DNA. Genes for
fluorescent proteins have been
inserted into their genome to produce
their fluorescent colors
 The human genome is composed of about 3 billion
base pairs of As and Ts, Gs and Cs. In 1990, an
international consortium of scientists set out to
create a map that would show exactly where on
our twenty-three pairs of chromosomes every one
of those base pairs is located. The effort, called
the Human Genome Project, is the most extensive
scientific enterprise ever undertaken.
 Genetic mapping is the first step in isolating a
gene. There are two basic approaches to this
complex endeavor. The first is called linkage
mapping, and its goal is to show where on each
chromosome each gene is located relative to
other genes. The method is to compare the
genetic makeup of members of a family or closely
related group of people who have a history of a
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