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BY:

DR. JAMILAH SYAFAWATI BINTI YAACOB, PhD


(Tel: 03-79674090, Fax: 03-79674178)

INSTITUTE OF BIOLOGICAL SCIENCES,


FACULTY OF SCIENCE,
UNIVERSITY OF MALAYA,
50603 KUALA LUMPUR,
MALAYSIA.
INTRODUCTION TO PLANT GENETIC
ENGINEERING

(CONCEPT OF INTEGRATION & EXPRESSION


OF ALIEN GENES IN TRANSGENIC PLANTS)
Genetic engineering (GE)

Definition: The modification of an organisms


genetic composition by artificial means, often
involving the transfer of specific traits, or genes,
from one organism into a plant or animal of an
entirely different species.

When gene transfer occurs, the resulting


organism is called transgenic or a GMO
(genetically modified organism).
Genetic engineering is different from
traditional cross breeding!

In cross breeding, genes can only be


exchanged between closely related species.

With genetic engineering, genes from


completely different species can be inserted
into one another.
Genetically modified crops first became
commercialized in mid-1990s.

Since then, their use has progressively


increased. In 2009, approx. 25% of all
agricultural crops were transgenic.
For example, scientists in
Taiwan have successfully
inserted jellyfish genes into
pigs in order to make them
glow in the dark.

In the US, there is a project


called Glowing Plant Project,
to produce natural lighting
(from plant) and save
electricity.
All life is made up of one or more cells. Each cell
contains a nucleus, and inside each nucleus are strings
of molecules called DNA (deoxyribonucleic acid).

Each strand of DNA is divided into small sections called


genes. These genes contain a unique set of instructions
that determine how the organism grows, develops,
looks, and lives.
During genetic engineering processes, specific
genes are removed from one organism and inserted
into another plant or animal, thus transferring
specific traits.

Eg:
(1) Bt corn gene from Bacillus
thuringiensis, and the gene of interest
produces a protein that kills Lepidoptera
larvae, (European corn borer)

(2) Bt cotton,

(3) Flavr Savr tomatoes, etc.


Transgenic soybeans from
Monsanto company,
resistant to glyphosate
(active agent in herbicide
Roundup). The herbicide
remains effective against
weeds, but not killing
herbicide-resistant crops.
Flavr Savr tomato developed by Calgene Inc. The plant was
given a gene that encodes an antisense RNA complementary to
the mRNA that encodes polygalacturonase enzyme. The
enzyme digests sugar linkaes within the pectin in cell walls of
tomato, hence softens the tomato.

Flavr Savr tomatoes have


longer shelf-life and
enhanced flavour.
CONCEPT OF INTEGRATION &
EXPRESSION OF ALIEN GENES IN
TRANSGENIC PLANTS
The production of transgenic plants is somewhat easier than
animals, because some somatic cells are TOTIPOTENT.

TOTIPOTENT: they are capable of developing into an entire


organism.

Hence, a transgenic plant can be made by introduction of


cloned genes into somatic tissue (eg. leaf tissue). Then, as the
cells of the leaf became transgenic, it can be treated with plant
growth hormones (through tissue culture), causing it to form
roots and shoots and subsequently into a complete plant.
TRADITIONAL SELECTION

Involves a long
process

Mostly genetically
blind

Limited by species
barrier
A transgene is a gene or genetic material that has been transferred
naturally, or by any of a number of GE or GM techniques from one
organism to another.

The introduction of a transgene has the potential to change the


phenotype of an organism.
Key step in cloning: cut & paste
of DNA fragments, to insert
chromosomal DNA into a
plasmid or viral vector.
Enzymes are used to cut DNA.
Called as restriction
endonucleases or restriction
enzymes (RE).
RE binds to a specific base
sequence and cleave DNA
backbone at 2 defined
locations (one in each strand).
Action is palindromic.
*Palindrome: a word or phrase that reads
the same backward as forward
To produce a recombinant DNA, both DNA from 2 different
sources (or plasmid) are cut with the same RE.
RE digests DNA into fragments with sticky end or blunt
end. Sticky ends are important in cloning as they allow
single-stranded regions of DNA to form hydrogen bond
to a complementary sequence of DNA (from different
source).

The hydrogen bonding between sticky ends promotes


temporary interaction between the two DNA fragments.

The interaction is not stable. Why?? Because it involves a


few hydrogen bonds only between the two fragments.

How to make it permanent? The sugar-phosphate


backbones within the DNA must be linked together by a
covalent bond. Linkage is facilitated by DNA ligase.

Video: Recombination
Gene cloning involves the insertion of DNA
fragments into vectors, which are then
propagated within host cells.

Video: Gene cloning


-galactosidase
Cleaves X-Gal
into a blue dye
Plant Biotechnology: The Genetic
Manipulation of Plants, by Adrian Slater,
Nigel W. Scott & Mark R. Fowler. Publisher:
Oxford University Press.

Chapter 18 & 19, Genetics: Analysis &


Principles, by Robert J. Brooker. Publisher:
McGraw Hill.