Teknologi Transfer Gen

Disusun oleh

Endang Semiarti
Fakultas Biologi UGM
2022

E. Semiarti_2022
BIOTEKNOLOGI MODERN→ Genetically Modified Organisms
Genetically Engineered Organisms

GEO
BIOCROPS
GMO

Fenotip dikode DNA

Rekayasa
Genetika

Semua makhluk hidup

tersusun atas DNA

E. Semiarti_2022
A Transgenic organism

Is an organism that has one or more genes that have

been artificially inserted
• These transferred genes are called transgenes.
-From a related or unrelated organism
-From a completely different genus
• Organism(s) that acquire new genes through the
process of crossing over are not considered to be
transgenic organisms.

E. Semiarti_2022
For example, transgenic
corn contains the Bt
gene from Bacillus
thuringiensis to control
the corn ear worm.
Over 80 million acres
planted annually saving
up to $2.4 billion in
pesticide use

http://plymouth.ces.state.nc.us/pub
s/btcorn99.html

E. Semiarti_2022
E. Semiarti_2022
E. Semiarti_2022
E. Semiarti_2022
Manipulasi Genetik pada Hewan

E. Semiarti_2022
E. Semiarti_2022
E. Semiarti_2022
E. Semiarti_2022
E. Semiarti_2022
E. Semiarti_2022
E. Semiarti_2022
E. Semiarti_2022
E. Semiarti_2022
E. Semiarti_2022
E. Semiarti_2022
E. Semiarti_2022
E. Semiarti_2022
E. Semiarti_2022
E. Semiarti_2022
E. Semiarti_2022
E. Semiarti_2022
E. Semiarti_2022
E. Semiarti_2022
E. Semiarti_2022
E. Semiarti_2022
 Plant
Biotechnology

Methods of
Plant Breeding

E. Semiarti_2022
How Can Genetically Engineered Plants Be Used?

•Agriculture
•Horticulture
•Forestry
•Environment
•Food Quality

E. Semiarti_2022
How Does Agrobacterium Gene Transfer
Work?

• Extract DNA from donor

• Cut DNA into fragments
• Sort DNA fragments
• Recombine DNA
fragments
• Transfer plasmids with
bonded DNA
• Grow transformed
(recipient) cells

E. Semiarti_2022
What do we need to be able to remove a
gene from one organism and insert it into
an other?

E. Semiarti_2022
We need
•A way to cut the DNA containing the gene
from the rest of the DNA
•A way to combine the gene with the DNA of
the recipient
•A way to insert the modified DNA into the
recipient
•Also, a way to read the bases

E. Semiarti_2022
Restriction enzymes

• Cuts DNA at a specific

sequence of 4-6 base
pairs
• The exposed bases
want to base pair with
complimentary “sticky
ends”

E. Semiarti_2022
Polymerase chain
reaction (PCR)
• Amplifikasi fragmen DNA
secara in vitro tanpa sel
• Bahan: DNA template,
DNA polymerase, dNTPs,,
primer/oligonukleotida
• Aplikasi:
psintesis/perbanyakan
DNA fosil, forensik,
prenatal diagnosis, dll.

E. Semiarti_2022
 Plasmids

•Small circular DNA

strands in some
bacteria

E. Semiarti_2022
Plasmids
•If plasmids are cut
with the same
restriction enzyme
as the gene of
interest the gene
and the plasmid will
have complimentary
sticky ends

E. Semiarti_2022
Gene insertion
•Many bacteria have the ability to take in
plasmids
•Just grow bacteria in a medium with lots of
plasmids
•Use a glass needle or a micro-gun to inject
DNA into eukaryotic cells
•Put the gene into a virus and hope the virus
“infects” the target cell

E. Semiarti_2022
Gene insertion

E. Semiarti_2022
 Transgenic Organisms

Organisms that contain foreign genes

E. Semiarti_2022
Transgenic bacteria

•Used to produce
human proteins
such as human
growth hormone,
insulin, and
interferon

E. Semiarti_2022
 Plant Transformation Methods

•Biological: Agrobacterium-mediated
Bacteriophage

•Non biological: Uptake of naked DNA

electroporation
particle gun
PEG
E. Semiarti_2022
Metode transformasi
Monokotil Dikotil
Mikroinjeksi + +
Transfeksi 0 +
(protoplas, Ca2+)
Elektroporasi 0 +
(protoplas)
Biolistic ++ +
T-DNA + ++

E. Semiarti_2022
Animal Transformation Methods

• Microinjection

• Direct transfer

E. Semiarti_2022
Membuat GM Plant melalui Agrobacterium

E. Semiarti_2022
Agrobacterium-mediated transformation

E. Semiarti_2022
Agrobacterium transformation

E. Semiarti_2022
Crown Gall disease

E. Semiarti_2022
E. Semiarti_2022
 Metoda Transformasi
protokorm
3 mg stlh tanam

Penaburan biji pada medium NP

Prekultur pd medium CIM,
4 hari
Phenotypic & PCR analysis

Ko-kultivasi
Transformasi 4 hari

menggunakan
Regenerasi A. tumefaciens
transformant

E. Semiarti_2022 Seleksi kandidat transforman Eliminasi bakteri

Plant Transformation

E. Semiarti_2022
 GM-Plant
Ornamental Food Plant

E. Semiarti_2022
Blue Rose

E. Semiarti_2022
 The expression of KNAT1 gene in the
black orchid transformants→multishoots

Bar: 1 cm
Black orchid transformants. A. Two-months old candidate of transformants grow on
kanamycin antibiotic containing medium in selection plate; B. A shoot grows from a
non-transformant protocorm; C. Multishoots grow from one 35S::KNAT1 transformant
protocorm;
E. Semiarti_2022
PENYISIPAN GEN PENGKODE ENZIM
LUCIFERASE

Kunang-kunang

Luciferase
E. Semiarti_2022
Tembakau berfluoresen
APLIKASI BIOTEKNOLOGI

Biopharmaceutical

Kesehatan

Hobi

E. Semiarti_2022
PRODUK REKAYASA GENETIKA
TUMBUHAN
Dengan rekayasa genetika, tumbuhan dapat digunakan untuk
memproduksi:

• Antibodi mamalia
• Vaksin
• Hormon
• Enzim
• Berbagai macam agen terapeutik

E. Semiarti_2022
 KELEBIHAN BUAH-BUAHAN DAN
SAYURAN
• Dapat dikonsumsi mentah atau olahan
• Sangat sesuai untuk produksi:
• Subunit vaksin
• Food additives
• Antibodi

Kentang Antibodi
Tomat Vaksin rabies pertama
(MeGarvey, 1995)
Selada Vaksin
(Kapustra et al., 1999)
Pisang
E. Semiarti_2022
Vaksin (Sala et al., 2003)
KELEBIHAN TANAMAN DALAM
BIOPHARMACEUTICAL

Tanaman mempunyai banyak kelebihan dibanding pembuatan

produk farmasi secara tradisional, karena:
• Biaya produksi rendah
• Perbanyakan cepat
• Bebas dari mikrobia patogen
• Produk protein yang dihasilkan persis sama dengan yang “ditiru”

E. Semiarti_2022
 Transgenic animals

• Animals that produce

omega-3 fatty acids

E. Semiarti_2022
Ikan hias transgenik

E. Semiarti_2022
 Animal transgenic

Glofish Transgenik

Andi
Polly dgn gen c-protein dari manusia

Neti & Dito

Sapi perah dgn gen-gen protein air

Babi mengandung gen manusia susu manusia

E. Semiarti_2022
@Budi Setiadi Daryono-2007
Stem sel

E. Semiarti_2022
Terapi gen

E. Semiarti_2022
4. BIOETHICS

E. Semiarti_2022
4. BIOETHICS
Bio-etika : Perilaku yang menghargai (respect)
hidup, mahluk hidup dan lingkungan hidup
berdasarkan norma-norma sosial - budaya dan
kehidupan berkeimanan

Bio-etika: Perlu dikembangkan untuk menjamin

keamanan dan keberlanjutan prinsip-pronsip
kehidupan, serta menjamin kesejahteraan umat

E. Semiarti_2022
BIOTEKNOLOGI
MODERN

E. Semiarti_2022
Referensi
Alberts, B., Johnson A., Lewis, J., Raff, M., Roberts, K., Walter, P., 2015,
Molecular Biology of the Cell. 6th ed. Garland Publ. Inc., New York.

Link website : Cell biology by Bruce #Alberts:

https://drive.google.com/file/d/1Ok_oiLldrtAo-
Y8eAKNKzBFo8reQCarN/view?usp=sharing

Win Darmanto (2019). Metode Transfer Gen Dalam Sel Mamalia.

https://slideplayer.info/slide/3209475/

Murni, D. (20`8). Hewan Transgenik

https://www.academia.edu/6341930/HEWAN_TRANSGENIK

E. Semiarti_2022