Tradition of Excellence

Production of Peptide Antihypertensives in the

Endosperm of Japonica Rice (Oryza sativa) Seeds

Cici Rizky Yonanda

JEMBER UNIVERSITY
Introduction
Tradition of Excellence

Hypertention is one cause

of premature death on
society in the world, WHO
has estimated by the year
2025 as many 1.5 billion
people in the world will
suffer from hypertention
every year. One of the
enzymes that play a role in
the occurrence of high
blood pressure are the
Angiotensin Converting
Protein seeds of the melinjo (Gnetum gnemon) hydrolyzed capable of
Enzyme (ACE).
lowering blood pressure, this is due to the activity of the ACE inhibitors that
are contained in the protein of seeds of melinjo hydrolyzed (Puspitaningrum
et al, 2014)
Research Scheme

2. Transformation
Tradition of Excellence
1. Construction and ligation

Preparing callus (Japonica rice)

Transformation to E. coli Transformation to E. coli

Gene ransformation by Agrobacterium tumefaciens into the callus of Japonica rice

Construction and ligation gene gG-7pAH with pGreen plasmid Co-cultivation

Elimination of Agrobacterium and callus selection in selection

Transformation recombinant plasmid to E. coli
medium

Putative transfoman and Aklimatisation

Recombinant plasmid verification with PCR

Plant characterization
Isolation DNA plasmid, cutting, dan sekuensing
Plasmid Construct
Tradition of Excellence

plasmid pGreenII 0229 10xPeptide. LB : Left border, RB : Right

Border, nos: nopaline synthetase gene, Glu4 Ter : Gluteline 4
Terminator, 10xPeptide Gnetum gnemon, OsGlu4 Pro : Gluteline
4 Promoter.
The expression of Gluteline Terminator and Gluteline Promoter
are directly into the endosperm of Japonica rice seeds.

Plasmid base
Transformation recombinant Tradition of Excellence
plasmid to E. coli
Recombinant plasmid
verification with PCR

M C1 C2 C3 C4 C5 C6

300bp
290bp

M (Marker); C (Colony of Bacteria)

Preparing callus (Japonica rice)
Tradition of Excellence

• Day 1 • Day 2
• Day 4

• Day 9 • Day 8 • Day 6

 Tradition of Excellence

Callus appearance with microscope at a weeks old

About 2 weeks old callus Tradition of Excellence
transfered to regeneration
medium
 First selection Second selection Third selection
Tradition of Excellence

Gene
ransformation by
Agrobacterium
tumefaciens into
the callus of
Japonica rice
Fourth selection Fifth selection Selection Percentage of living
callus
Tradition of Excellence
1 100%

2 80%

3 60%

4 50%

5 45%
Tradition of Excellence