Kuliah Bioteknologi

Jurusan Biologi FMIPA

Pengampu :
1. Bambang Sugiharto (Prof. Dr.)
2. Esti Utarti (SP., MSi)
3. Sattya Arimurti (SP., MSi)
Materi :
1. Kuliah
2. Praktikum

Kuliah
1. Pendahuluan, pengertian,
perkembangan dan ruang lingkup
2. Genom DNA
3. Ekspresi gen
4. Regulasi ekspresi gen
5. Transformasi DNA
6. Rekombinasi DNA
7. Pustaka Genom

Bioteknologi diartikan sebagai pemanfaatan

mikroorganisma untuk menghasilkan produk
yang bermanfaat
Awal pemanfaatan bioteknologi meliputi :
Fermentasi juice untuk minuman (jaman prasejarah)
Fabrikasi pembuatan wine (abad 3)
Pembuatan tape dan tempe (???)
Produksi ethanol dan vinegar (abad 12-14)
Penemuan yeast dan fermentasi asam laktat (abad 19)
Penemuan Pinicilin (1928)
Penemuan senyawa antibiotik (diawali sekitar 1945)

Pengembangan tehnik isolasi mikroba, perbaikan

sifat organisma dengan mutagenesis,
perkembangan penemuan DNA dan tehnik
rekombinan DNA telah memacu perkembangan
bioteknologi modern.
1953 Double helix structure of DNA is first described by Watson
and Crick.1973 Cohen and Boyer develop genetic engineering
techniques to "cut and paste" DNA and to amplify the new DNA in
bacteria.1977 The first human protein (somatostatin) is produced
in a bacterium (E. coli).1982 The first recombinant protein
(human insulin) appears on the market.1983 Polymerase chain
reaction (PCR) technique 2000 A first draft of the human genome
sequence is completed.2005 Over 40 million gene sequences are
in GenBank, and genome sequences of hundreds of prokaryotes
and dozens of eukaryotes are finished or in draft stage

More food
will be needed
to feed a
growing global
middle class

Developed world
(EU, U.S., Japan)
Population: 1 billion
Income: $5,000+
Developing world
(Asia, Latin America)
Population: 4.2 billion
Income: $400 $5,000

Impoverished areas
(Africa)
Population: 800
million
Income: <$400
Benefits of biotechnology More food

By 2025, there will

be another 2 billion
mouths to feed
United Nations
Population Fund

Benefits of biotechnology More food

Selective
breeding led to
higher-yielding
varieties.

What is plant biotechnology?

Traditional plant breeding

DNA is a strand of genes,
much like a strand of
pearls. Traditional plant
breeding combines many
genes at once.

Traditional donor

Commercial variety

New variety

(many genes are transferred)

X
(crosses)

Desired Gene

Desired gene

Plant biotechnology
Using plant biotechnology,
a single gene may be
added to the strand.

Desired gene

Commercial variety New variety

(only desired gene is transferred)

=
(transfers)
Desired gene

What is plant biotechnology?

Plant biotechnology definition:

A precise process in
which scientific
techniques are used to
develop useful
and beneficial plants.

What is plant biotechnology?

Benefits of
biotechnology
More food
Better
food

Better for the

environment

Four countries accounted for 99 percent* of

the global biotech crop area in 2002

*Australia, Bulgaria, Colombia, Germany, Honduras, India, Indonesia, Mexico, Romania, South
Africa, Spain and Uruguay accounted for the remaining 1 percent of biotech crop acres.
Source: International Service for the Acquisition of Agri-biotech Applications

Products on the market

Global plantings of biotech crops

increased by 12 percent in 2002

Million Hectares

Global Area of GM Crops

Source: International Service for the Acquisition of Agri-biotech Applications

Products on the market

SUGARCANE INDUSTRY IN INDONESIA

Had been started from
Holland Gov.
Recent variety - low
sugar productivity
Development
biotechnology for
sugarcane

CARBON ASSIMILATION AND SUCROSE

BIOSYNTHESIS IN SUGARCANE

Phosphoenolpyruvate
carboxylase (PEPC)
Sucrose-phosphate
synthase (SPS)

r = 0,89
3

0,8

2
7
1

0,7
4

0,5

0,55

0,70

0,85

Plant Growth (gram/day)

r = 0,92

6
7

0,9
2

0,8

0,7
1

0,6
4

0,5

40

50

60

Leaves Sucrose (mM/gram Fw)

SPS Activity (mM/gram Fw)

1,0

1,0
SPS Aktivity (mM/gram Fw)

SPS Activity (mM sucrose/gram Fw)

Sucrose-phosphate synthase activity

determines growth and sugar productivity in
sugarcane
0,9

r = 0,81

2
7

0,8
8

3
1

0,7
0,6

6
9
12
Sugar productivity (% Brix)

Correlation between SPS activities and plant growth (left), leaf sucrose content
(middle) and sugar productivity (right) in sugarcane.

Source : Sugiharto et al., Zuriat 7:76-85 (1996); Sugiharto et al., Proceeding

of Xth International Congress on Photosynthesis, France (1995)

Relative levels of mRNAs

Isolation of two genes for SPS from

sugarcane that differentially expressed
10
8
6
4
2
0

SoSPS1

12

SoSPS 2

24

48

Time after illumination (h)

Comparison of the amino acid
sequences deduced from cDNA for
SPS from sugarcane and from
others sources (left)
Differential expression SoSPS1 and
SoSPS2 in sugarcane leaf under
illumination (upper)

Source : Sugiharto et al., Plant

Cell Physiol. 38:961-965 (1997)

Transformation of sugarcane
SoSPS1 gene in tobacco
600
kb

P3
NOSPro

NPT II (Kan R)

NOS-Ter

P1

Pro CaMV

SPS tebu
P4

700
kb

Transgenic tobacco
expressed SoSPS1 gene

P2

NOS-Ter

Construct of SoSPS1
in modified pBI121

Over-ekspresi gen SoSps1 pada

tanaman tebu (proyek RUT VIII)
Xba1
pTopoSPS

BamH1 Xba1BamH1
SPS1

Xba1
NP NPTII NT CaMV35S

SPS1

Transformation into
Agrobacterium cell

BamH1
NT

pB1121-SPS1 construct

Infected to
sugarcane callus
Outline the method to
create transgenic
sugarcane

Source :

Regeneration of
transformant

Sugiharto et al., RUT

VIII Report (2003)
Transgenic cane