Mekanisme Transfer Gen

secara
in vitro dan in vivo

Disusun oleh
Endang Semiarti
Fakultas Biologi UGM
2021

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BIOTEKNOLOGI MODERN→ Genetically Modified Organisms
Genetically Engineered Organisms
GEO
BIOCROPS
GMO

Fenotip dikode DNA

Rekayasa
Genetika

Semua makhluk hidup

tersusun atas DNA

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 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.

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For example, transgenic
corn contains the Bt
gene from Bacillus
thuringiensis to control
the corn ear worm.
→ Bt Toxin production in
plant
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
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Manipulasi Genetik pada Hewan

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 Plant
Biotechnology

Methods of
Plant Breeding

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How Can Genetically Engineered Plants Be Used?

•Agriculture
•Horticulture
•Forestry
•Environment
•Food Quality

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 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

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

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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

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Restriction enzymes

• Cuts DNA at a specific

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

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 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.
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Plasmids

• Small circular DNA

strands in some
bacteria

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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

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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

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Gene insertion

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 Plant Transformation Methods

•Biological: Agrobacterium-mediated
Bacteriophage

•Non biological: Uptake of naked DNA

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

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Animal Transformation Methods

• Microinjection

• Direct transfer

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Membuat GM Plant melalui Agrobacterium

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Mechanism of Agrobacterium-mediated
transformation

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Agrobacterium-mediated transformation

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Crown Gall disease

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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

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Seleksi kandidat transforman Eliminasi bakteri
Plant Transformation

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GM-Plant
Ornamental Food Plant

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Blue Rose

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 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;

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PENYISIPAN GEN PENGKODE
ENZIM LUCIFERASE

Kunang-kunang

Luciferase Tembakau berfluoresen

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APLIKASI BIOTEKNOLOGI

Biopharmaceutical

Kesehatan

Hobi

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

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

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KELEBIHAN BUAH-BUAHAN DAN
SAYURAN
• Dapat dikonsumsi mentah atau olahan
• Sangat sesuai untuk produksi:
• Subunit vaksin
• Food additives
• Antibodi
Antibodi
Kentang
Vaksin rabies pertama
Tomat
(MeGarvey, 1995)
Vaksin
Selada
(Kapustra et al., 1999)
Vaksin (Sala et al., 2003)
Pisang
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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”

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Transgenic animals

•Film “ The mouse

that Laid the
Golden Egg”
•Pigs that produce
omega-3 fatty acids

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Ikan hias transgenik

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 Animal transgenic

Andi
Glofish Transgenik

Polly dgn gen c-protein dari manusia

Neti & Dito

Sapi perah dgn gen-gen protein air

Babi mengandung gen manusia
susu manusia

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@Budi Setiadi Daryono-2007
Stem sel

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Terapi gen

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4. BIOETHICS

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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

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BIOTEKNOLOGI
MODERN

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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

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