GENETICALLY

ENGINEERED CELLS
Genetic Engineering
genetic modification
The direct manipulation of an
organisms genome using
biotechnology
Genetically Modified Organism
(GMO) an organism generated
through genetic engineering
History of Genetic
Engineering
1973- first GMOs were bacteria
1974-genetically modified mice
1982-commercialized insulin-
producing bacteria
1994-genetically modified food
CLONING
Biology vs. Biotechnology
Cloning is the Cloning is the
process of
producing similar processes used to
populations of create copies of
genetically identical 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.
CLONING
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.

Natural Cloning
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.
Natural Cloning
Some animals, clone themselves by
parthenogenesis e.g. Little Fire Ant while
some clone themselves via fragmentation,
e.g. starfish
Molecular Cloning
Cloning of DNA fragments involves four
steps:
1. Fragmentation
2. Ligation
3. Transfection
4. Screening/selection
Molecular Cloning
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
Genetic Engineering of Higher
Organisms

Transfection the process of introducing

foreign DNA into eukaryotic cells,
introduction of foreign gene into higher
organisms.
Genetic Engineering of 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
 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.
 Genetic Engineering of Higher
Organisms
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.
 Genetic Engineering of Higher
Organisms
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.
Genetic Engineering of Higher
Organisms
 Genetic Engineering of Higher
Organisms
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.
Genetic Engineering of Higher
Organisms
GENE INSERTION
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
GENE INSERTION
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.
GENE INSERTION
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
GENE INSERTION
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