Genetic Engineering

genetic modification
The direct manipulation of an
organisms genome using
biotechnology
Genetically Modified Organism
(GMO) an organism generated
through genetic engineering
1973- first GMOs were bacteria
1974-genetically modified mice
1982-commercialized insulin-
producing bacteria
1994-genetically modified food
 Biology vs. Biotechnology
Cloning is the Cloning is the
process of
producing similar processes used
populations of to create copies
genetically identical of DNA
individuals that
fragments(molec
occurs in nature
when organisms ular cloning),
such as bacteria, cells (cell cloning)
insects or plants or
reproduce asexually
microorganisms.
 Natural Cloning-It is the reproduction
method used by plants, fungi and bacteria
the way that clonal colonies reproduce
themselves.
e.g. blueberry plants, hazel trees
Molecular Cloning- It refers to the process
of making multiple molecules.
 single-celled organisms, clone themselves via
binary fission
e.g. paramecium, bacteria, many algae

Plants, vegetative reproduction often involves

the formation of adventitious roots or
shoots.
 Some animals, clone themselves by
parthenogenesis e.g. Little Fire Ant while
some clone themselves via fragmentation,
e.g. starfish
 Cloning of DNA fragments involves four
steps:
1. Fragmentation
2. Ligation
3. Transfection
4. Screening/selection
 Fragmentation breaking apart a strand of
DNA
Ligation- gluing together pieces of DNA in a
desired sequence
Transfection- inserting the newly formed
pieces of DNA into cells
Screening/selection- selecting out the cells
that were successfully transfected with the
new DNA
 Transfection the process of introducing
foreign DNA into eukaryotic cells,
introduction of foreign gene into higher
organisms.
 Some plants are subject to infection with the
bacterium Agrobacterium tumefaciens
contains a plasmid that contains a section
known as T-DNA. This T-DNA can integrate
into the plant chromosome. If genes are
inserted into the T-DNA region, they can be
incorporated ultimately in the plant
chromosome. Unfortunately, most cereal
plants are not readily susceptible to
Agrobacterium
 Genetic Engineering of Higher
Organisms
Biolistic Process
It is a new process which employs high
velocity microprojectiles to deliver substances
into cells and tissues. The biolistic process
represents a completely new approach to the
problem of how to deliver DNA into intact cells
and tissues. High velocity microprojectiles are
used to carry DNA or other substances past cell
walls and membranes.
 Electroporation
It is a microbiology technique in which an
electrical field is applied to cells in order to
increase the permeability of the cell membrane,
allowing chemicals, drugs, or DNA to be
introduced into the cell. In microbiology, the
process of electroporation is often used to
transform bacteria, yeast, or plant protoplasts
by introducing new coding DNA.
 Protoplasts
In biology, it was proposed by Hanstein
(1880) to refer to the entire cell, excluding the
cell wall.
Protoplasts also refer to a plant, bacterial
or fungal cell that had its cell wall completely or
partially removed using either mechanical or
enzymatic means.
 Baculoviruses
For most animal cells, genetic
manipulation can be accomplished by
modifying viruses to become vectors. For
example, in the insect cell system a baculovirus
can be modified so as to place a desired gene
under the control of a very strong promoter at
the expense of a gene product that is
unessential for viral replication in cell culture.
Insertion of Genes into Plants
Crop plants
Addtion of gene may improve the
agricultural, hotricultural or ornamental
value of crop.
Transgenic plants can act as living
bioreactors for the inexpensive
production of economically proteins or
metablites.
Plant genetic transformation
(transgenesis) provides a powerful
means for studying the action of genes
during development and other processes
Insertion of Genes into Animals
The production of transgenic animals has
focused mainly on producing models, for
instance in the mouse for basic and medical
research.
Transgenic chickens could be used to improve
the genetic make-up of existing strains with
respect to built-in resistance to viral, bacterial
and coccidial disease, better feed efficiency,
lower fat and cholesterol levels in eggs and
better meat quality.
Insertion of Genes into Animals
Transgenic fish-transgenes have been
introduced by microinjection or
electroporation of DNA into the fertilized
eggs of a number of fish species including
carp, catfish, trout, salmon, Arctic char and
tilapia
Insertion of Genes into Microorganisms
A high number of bacteria and yeasts have
been genetically engineered for production of
industrially, nutritionally and medically
important eukaryotic gene products under
contained conditions. e.g. Saccharomyces
cerevisiae and Pichia pastoris