BTY100-Lec#5.

1
Genetic Engineering

LPU : BTY100

Outline
Introduction to Genetic Engineering
Goal
Tools
Molecular Scissors
Cloning Vectors

Process
Enhanced Traits- Bt Crops

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Genetic Engineering
Altering/ Manipulating the traits of living
organisms by changing the information
encoded in their DNA.

Goal
To add one or more new traits that are not
naturally already found in that organism.
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Altering DNA? Is it possible?

DNA is a universal language.
The genetic code means the
same thing in all organisms.
All living organisms have the
same bases (A, T, G, C) which
code for proteins.
The codons they form always
code for the same amino acids,
so transferred DNA codes for
the same polypeptide chain in
the host organism as it did in
the donor organism.
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Human
appearance is a results of the
information stored in our genes. Differences in
Eye color, hair color, skin complexion, height etc
all are due to different information stored in
different persons.
Does it means any change in our DNA will
result in some change in our appearance?

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Changing faces of Michael

A Question
for you.?
Jackson.?????
In one of the most memorable moments of Oprah's interview, Michael
told her he had a skin disorder that destroyed the pigmentation of his
skin. The disease, called vitiligo, was in his family, Michael said. "It is
something I cannot help.

Genetic
Engineering?
OR
Skin Bleaching?
OR
Skin Disease???

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He kept getting whiter

and whiter and whiter,
and nobody understood
why

Coming back to our Question

Does it means any change in our DNA will
result in some change in our appearance?
AnswerNO, The Purpose of science is not
this. Ultimate aim of all type of researches is
to benefit the society.
In this unit we will be discussing the examples
which were created for some useful purpose.
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How does Genetic Engineering

work?
removing a gene from one organism and inserting
it into another (a process called gene transfer).
When a gene for a desirable trait is taken from one
organism and inserted into another, it gives the
recipient organism the ability to express that
same trait.

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Glossary
Donor cell- is the cell that provides the new DNA
fragment for the recipient cell
Recipient cell/Host Cell- is the cell that is
reviewing the new DNA.
Target Gene/Foreign DNA- Gene with the desired
trait.
Ligation- Uniting attaching two DNA fragments.
DNA recombination- The DNA fragment to be
cloned is inserted into a vector.
Transformation- The recombinant DNA enters into
the host cell and proliferates
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GENETIC ENGINEERING:
PROCESS

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PROCESS:

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Tools for Genetic Engineering

Molecular Scissors
Construct/Cloning Vectors

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Molecular Scissors
Restriction
enzymes
areDNAcutting enzymesfound in bacteria.
A restriction endonuclease : cuts
DNA at or near a particular place
known as restriction sites.
E.g. EcoRI:
always cuts at site: GAATTC

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Cloning
Vectors
A DNA molecule that:

carries foreign DNA into a host cell.

replicates inside cell and produces many copies of itself and the foreign
DNA.

E.g.: Plasmid: an extrachromosomal circular DNA molecule that

replicates on its own inside the bacterial cell

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Transformation
The plasmids carrying the
desired traits are introduced
into host cells via a process
called transformation.
Bacteria are most commonly
used as host cells for gene
cloning because :
1.Their DNA can be easily
isolated and reintroduced into
their cells.
2.Bacterial cultures also grow
quickly, rapidly replicating the
foreign
genes.
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Process

Gene of interest is cut out with Restriction enzyme.

Host plasmid is cut with same Restriction enzyme.

Gene is inserted into plasmid and ligated with

ligase enzyme

New plasmid inserted into bacterium (transform).

Bacteria divides and plasmid divides along with

that.

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Genetic Engineering:
Enhanced Traits

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Bt Crops
Are crops that are genetically engineered to produce the same
toxin as produced by B. thuringiensis in every cell of the plant,
with the goal of protecting the crop from pests.
Examples: Bt Cotton, Bt corn, Bt Brinjal

What isBt?
Bacillus
thuringiensis(Bt
) is a spore
formingbacteriu
m that produces
crystals protein
(cry proteins),
which are toxic
to many species
of insects
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Bt Cotton
Bt Cotton is a variety of cotton that has been genetically
engineered to produce an insecticide in every cell.
It contains the gene coding for Bt toxin, causing cotton plant to
produce this natural insecticide in its tissues.
In many regions, the main pests in commercial cotton are
lepidopteran larvae, which are killed by the Bt protein in the
transgenic cotton they eat.
This eliminates the need to use large amounts of broad-spectrum
insecticides to kill lepidopteran pests

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How Bt Works?
1. When insects attack and eat the
cotton plant the Cry toxins are
dissolved.
2. This is made possible due to the
high pH level of the insects
stomach.
3. The now dissolved and activated
Cry molecules bind to cells in the
insect gut and results in the death
of epithelial cells.
4. The death of such cells creates
pores in the insect gut and insect
dies eventually.
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Why Bt is not toxic to

Humans?
The specificity of Bt rests on the fact:
1. For the activation of toxin, alkaline conditions are required as provided by
insect gut but the human stomach has a pH of 1.5 to 3.5.
2. The toxicity of the Bt protein is receptor-mediated. This means that for an
insect to be affected by the Bt protein, it must have specific receptor sites
in its gut where the proteins can bind. Humans do not have these receptors.
Before Bt crops are placed on the market, they must pass very
stringent regulatory tests, including those for toxicity and
allergenicity.

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Advantages
Reduced environmental impacts from pesticides
Increased opportunity for beneficial insects
Reduced pesticide exposure to farm workers
and non-target organisms

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Risks
Invasiveness Genetic modifications can
potentially change the organism to become
invasive.
ResistancetoBt- The biggest potential risk to
usingBt-crops is resistance. Farmers have taken
many steps to help prevent resistance.
Cross-contamination of genes - Although
unproven, genes from GM crops can potentially
introduce the new genes to native species.
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Next Class:

Genetic
Engineering:A Potential
Solution

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