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GENETIC ENGINEERING

Presented by:
1. Wahyu Maulana Endris 15725251010
2. Royan Mahmud Musthofa
15725251038

PROGRAM STUDI PENDIDIKAN BIOLOGI


FAKULTAS PASCASARJANA
UNIVERSITAS NEGERI YOGYAKARTA
2016

1. HISTORICAL OF GENETIC ENGINERING


2. Recombinant dna technology
3. Base technology of dna recombinant
4. Equipment recombinant dna technology
5. Cellular enzyme

Contents

6. Natural vector
7. Benefit of technology recombinant
8. Techniques of Genetic Engineering
9. Steps of Genetic Engineering
10.Application of Genetic Engineering in
Daily Life
11.Bioethics

Wahyu maulana endris | 1572525101

1931

Phenomena recombination
and physical evidence DNA
was found.

1968

Endonuclease was discover or


enzyme restriction done by
stewart linn and werner arber.

1973
1. Historical
of genetic
enginering

The first experiment in


recombinant dna cloning
done by herbert boyer
and stanley cohen .

Wahyu maulana endris | 1572525101

1979
1980

First mice genetically


engineered was developed.

A method of
producing insulin
using genetic
engineering was
found .

1991

Gene therapy first tried


and tested in humans .

1994

Tangential tomatoes released in


the market for the first time

1. Historical
of genetic
enginering

1997

Dolly, the first animal


clone

Wahyu maulana endris | 1572525101

2.
Recombinant
DNA
Technology
A long time ago, our forefathers aware of
diversity living things.
Diversity this living thing has allowed humans to
choose which type of living things he directed .
Technique recombinant dna is a technique in
genetically engineered to produce new
properties by means of Recombining a particular
gene of the dna with the genome .

2.
Recombinant
DNA
Technology
Technology known as recombinant dna
technology, or with the term more
popular is genetic engineering , this
involving effort making copies of a
particular gene in a cell that is not its
natural cell buses often also was said to
be cloning gene.

3. Base
technology of
recombinant dna
Transfer dna or
displacement dna or
displacement dna into
bacteria can in three
ways, namely conjugation,
transformation, and
transduction.

Conjugation

Transformatio

Transductio

Conjugation

Resistant Bacterium

Sensitive
Resistant Bacterium

Transformation

Transduction

4. Sets of equipment for dna


recombinant technology
Restriction Enzyme

Ligase Enzyme

VECTOR
PLASMID
COSMID
BAKTERIOPHAGE

Transposon
Dna/rna detector
Genom map

5. Cellular
enzyme

Restriction
Endonuclease
DNA/RNA
polymerase
Dna ligase

5. Cellular
enzyme

reverse
transcriptase

6. Natural
Vector

An instrument that
carries dna into his
new stem cells.

7. Benefit of
technology
recombinant

Application technique
recombinant dna in
biotechnology of
them are vaccine
production , insulin ,
antibodies and forth.

Technique of DNA Recombinant

DNA Recombinant give us ability to take the specific genes


from organism, combine those gene to other organism,
insert to the host cell, until the ability of cell to express those
kind of combining gene.
There are several technique of DNA recombinant

DNA
Insertio
n

DNA
Isolation
DNA
Cutting
DNA
Combini
ng

1 DNA Isolation
Taking the DNA form the target cell (genomic DNA and
plasmid DNA)
Crushing the cell membrane or cell wall by mechanical
or enzymatically ways
Crushing the nucleus membrane by adding the Triton-X
100 or Sodium Dodecyl Sulphate (SDS)
Separating the DNA from other cell organelles by
sentrifugation
Purifying the DNA by adding the ammonium acetate or
alcohol

Diagram of DNA Isolation

2 DNA Cutting
Cutting the DNA by restriction endonuclease
enzyme
Endonuclease enzyme able to recognizing,
binding, and cutting the restriction site.
For example; EcoRI enzyme will cut the DNA which
specifically has the base sequence of GAATTC
The cutting process will result the asimetris point
(stickey end), meanwhile the enzyme which cut
the DNA in blunt (blunt end)

Diagram of DNA Cutting

Sticky End Vs. Blunt End

3 DNA Combining
The
important
condition
of
DNA
combining
is
the
genomic DNA must be
compatible with the
vector (plasmid) DNA
Three
ways
to
combine the genomic
DNA with the plasmid
DNA

Using the
DNA ligase
enzyme
from
bacteria

Giving
deoksinukleotidi
l
transferase
enzyme
to synthesize
single
strand of
homopolimeric

3
Using the DNA
ligase from E. Coli
which has
infected by
bacteriophage T4

Diagram of DNA Combining

4 DNA Insertion
DNA insertion also known as transformation, found by M.
Mandel dan A. Higa (1970)
The DNA recombinant and bacteria (host cell), mix in the
solution of CaCl2 in the temperature 0o 5oC
CaCl2 solution will make the bacteria cell to be swollen, make
the cell wall lose the periplasmic protein and finally leak
The recombinant DNA will bind with Ca2+, forming the DNAse
complex resistant
Those complex will finally enter the bacteria by heat-shock
treatment

Diagram of DNA Insertion

Transformant Selection
The selection of the cell which contain the vector which carry
the DNA recombinant can be detected using the tracefragment (probe)
The making of probe using Polymerase Chain Reaction (PCR)
by in vitro
The technique to detect the host cell contain desired gene
using colony hybridization.
The DNA of cell, sunk in the trace solution, so the tracefragment (probe) will insert to the DNA
Next, the position of DNA which inserted by the probe will
compare to the colony position of initial culture (master plate)

Because of not all cell (bacteria) contain the


recombinant DNA, so we must select the cell
that carry the recombinant DNA. There are 3
possibilities that can happen after
transformation:
The host cell didnt inserted by any
other gene
The host cell religation
The host cell inserted by recombinant
vector with or without the expected
gene

STEPS of DNA Recombination

(summary)

Isolation of DNA
Cutting DNA
Combining DNA
Analyze DNA
Transformant and recombinant
selection

ST
EP
S
of
D
NA
Re
co
m
bi
ati
on
(sum
mary)

Application of Genetic Engineering in Daily Life

The development of genetic engineering is


very advanced through the invention of
modifying genetic
The researcher has produced much more
product and transgenic organism by
genetic engineering known as Genetically
Modified Organism (GMO) or Organisme
Hasil Modifikasi Genetik (OHMG)

Many Sectors Have Affected by Genetic Engineering

1. Farms
Transgenic plant that;
Tolerant to chemical
substances
Resistant to diseases
Has the specific
characteristic
Able to bind N2 from
the air autonomously
Have a good
adaptation

2. Horticulture and
Forestry

3. Pharmacy and Industry


Making Insulin Through Genetic
Engineering
Isolation of Insulin Gene and Vector
1
Plasmid contain gene Amp-R and LacZ
2

Insert Insulin Gene into Vector

Cutting DNA which containing insulin and also plasmid


Combine the fragment DNA (Insulin) and plasmid
Use Ligase to covalently bind fragment and plasmid

Insert Plasmid Into Bacteria

Modified plasmid mix with the bacteria culture


Bacteria will take the plasmid through transformation

Cloning of Gene and Host Cell

The bacteria as the result of transformation are placed in the solid


medium which contain ampicillin and sugar (X-Gal)

Identify the Clone

Using the trace-fragment (probe) nucleid acid probe

Di
ag
ra
m
of
M
ak
in
g
In
su
lin

4. Environment

Some transgenic bacteria will help bioremediation of polluted coast and river

5. Laws and
Forensic

Bioethics
In the development of Science, the
researcher and technocrat whom plunge
into activity of molecular biology and
genetic engineering must giving attention
to the rules and ethics.
Bioethics giving the solution to solve the
controversy in the experiment,
development, and the usage of human
and natural resources


U
O
Y
K
N
A
H
T

bye bye.

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