Karnival Pendidikan

Melaka 2009

Biology Molecule
Toxicology
Chemistry & Food Technology
Plant Tissue Culture
Microbiology
 Melaka ‘Education Fair 2009’
Institute of
Biotechnology

Applications of Molecular Biology

Techniques in Biotechnology
Industry

Amelia Kassim
Nurhuda Jalil
Nurul Arneida Husin
 INTRODUCTION
Molecular biology is the study of
biology at a molecular level.

One of the most basic techniques of

molecular biology to study protein
function is expression cloning
Disciplines relation
Market intelligence

Plant Tissue
Culture
Melaka
InstituteProteomic
of Microbiology
Biotechnology Bioinformatic
 What is DNA cloning?
n When DNA is
extracted from an
organism, all its
genes are obtained


n In gene (DNA)
cloning a particular
gene is copied
(cloned)
 What is Gene cloning?

•To "clone a gene" is to

make many copies of it
•Act of making many
identical copies of gene
•Gene can be an exact
copy of a natural gene
•Gene can be an altered
version of a natural gene
 How is DNA cloned?

Cell-based DNA cloning Cell-free DNA cloning (PCR)

Clo n e in d iv id in g c e lls
 Clo n e in P CR
First cycle DNA sequence to be amplified

Denaturation,
Primer oligonucleotide annealing

DNA replication

Second cycles

20 – 30
cycles

Amplified DNA
sequences
Some Molecular Biology Techniques
1.DNA / RNA extraction
2.pDNA purification
3.Polymerase chain reaction (PCR)
4.Expression cloning
5.Gel electrophoresis
6.Gel documentation, and image analysis
7.Southern blotting
8.Northern blotting
9.Western blotting and immunochemistry
10.cDNA / genomic DNA library construction, and its screening
11.Southern, and Northern Hybridization techniques
12.Auto-radiography
13.Primer designing
14.PCR-techniques
15.PCR product Cloning
16.Construction of transformation vectors
17.Biolistic PDS He/1000 mediated plant transformation
18.Agrobacterium mediated genetic transformation
19.Analysis of the transgenic plants
20.Microscopy
21.Cultivation of bacteria / Microbiology,
22.preparation of glycerol stocks
23.Generation of ESTs
24.Analysis of DNA, cDNA / ESTs sequences using online and offline bioinformatics tools
25.Promoter isolation and characterization ++++++++++++++++++++++++++++++
Wh e n y o u a re w o rk in g in
Mo le c u la r Bio lo g y La b

Melaka
Institute of
Biotechnology
Gel-Documentation
RT-PCR
Spectrophotometer
 Gel Electrophoresis

Melaka
Institute of
Biotechnology
 Applications of Molecular Biology in
Biotechnology Industry ?
Some Examples:
1.Genetic engineering of plants / Animals/ Microbes
2.DNA Fingerprinting
3.Novel (Blue rose)
4.Exploitation of natural resources plant product
development (Genes for Perfumes in Plants)
5.Gene Cloning - Insulin and other drugs (HIV?) production?
6.Criminals monitoring based on DNA
7.Fruits with insulin, Milk with Antioxidants / vitamin A ++
Power of Metabolic Engineering

Metabolic Engineering
Genetically modified
 DNA
FINGERPRINTIN
G
TECHNOLOG
Y Prepared by:
Nurul Arneida Husin
Molecular Bioinformatics
 DNA???
•Deoxyribonucleic acids
•Building blocks of life.
•Nucleic acid that contains within it the genetic
code that makes up the blueprint of the
biological development of all plant, animal and
some virus life forms.
•Our unique marker that can uniquely identify
us.
•No two organism will ever have the same
DNA, except in rare cases of twins.
•
Deoxyribonucleic acid (DNA) is a nucleic acid that
contains the genetic instructions used in the
development and functioning of all known
living organisms and some viruses.
 By 1953,
Crick and Watson
had built a model which
incorporated all known
features of DNA, and
proposed the double helix
structure which is
commonly referred to as the
Watson-Crick model of
DNA.

Crick, Watson and Wilkins

were awarded Nobel prize
for this work in 1962.
BASE PAIRING

A-T
G-C
A- ADENINE
T-A
T- THYMINE
C-G
G- GUANINE
C-G
C- CYTOSINE
G-C
A-T
 DNA
FINGERPRINTING?????
•Known as DNA PROFILING.
•Techniques for analyzing and
comparing DNA from separate sources,
to identify any organism (human, plant,
animals, microbes etc) from their
biological materials.
 How to perform DNA fingerprinting?

•Study and Characterization of Plant Morphology

•Selection and Identification of Plants
•Isolation of Genomic DNA
•Selection of Specific Primers
•PCR (Polymerase Chain Reaction) Amplification
•Gel Electrophoresis Analysis
•DNA Analysis of Species-Specific Marker
•Development of Plant Database
 DNA Fingerprinting Process

Selection
of Primer/s

Raw materials
Adding PCR
Isolation of Component
Genomic DNA and tested
samples
Data analysis

Run Gel Electrophoresis

Run PCR
PCR product

Unique DNA Fingerprint profiles Loading PCR product

What is DNA Marker Technology?

DNA Marker Technology is a set of

technologies such as Random Amplified
Polymorphic DNA (RAPD), Amplified
Fragment Length Polymorphism (AFLP),
and Microsatellites.
 Application to herbal industry?
•DNA Marker Technology has found application in
fingerprinting genotypes, in determining seed
purity, in systematic sampling of germplasm, and
in phylogenetic analyses.
•DNA based techniques have been widely used
for authentication of plant and herbal species of
medicinal importance. This technology is very
helpful for the identification of the variety within
species or species within the generic group.
•Global trend leading to increased demands of
medicinal plants for pharmaceuticals,
phytochemicals, nutraceuticals, and cosmetics.
•Therefore, this service technology is applicable for
those in herbal business sector to make quality
assurance and correct identification of the starting
materials to ensure reproducible quality of herbal
medicine, which contributes to its safety and
efficacy.
•It is because herbal medicinal products may vary in
composition and properties, unlike conventional
pharmaceutical products, which are usually prepared
from synthetic, chemically pure materials by means
of reproducible manufacturing techniques and
procedures.
•This application of markers can be explored for
standardization of botanicals formulation containing
medicines.
Molecular Markers in Herbal Drug
Technology
•Herbal drug technology….is used for
converting botanical materials into
medicines, where standardization &
quality control with proper
integration of modern scientific
techniques & traditional knowledge
is important.
•
•
Roles of Molecular Markers in Herbal Drug
Technology?
•Authentication of plant species of medical
importance.
•Genotyping & quick identification of botanical
samples.
•Detection of adulteration/substitution
•Marker assisted selection of desirable
chemotypes
•Medicinal plant breeding
•Applications in foods & nutraceuticals
•
Molecular Markers in Essential Oils
Production
 Roles of Molecular Markers in
Increasing Essential Oil Production…..

•Breeding strategy
•To identify right cultivar for increasing
essential oils production.
•
•DNA Marker study will provide information on
ways of enhancing the economics of
essential oil production.
Hom
e
Microbiology World
and You

MICROBIOLOGY UNIT
Be one step ahead of the challenge
become a Microbiologist!
What is Microbiology?
◗ Micro - too small to be seen with the naked
eye
◗ Bio - life
◗ ology - study of
Why study Microbiology?
◗ Microbes make the world go round.
Microbiology lies at the heart of the
biosciences. There is hardly an area of
medicine unaffected by microbes and
microbes are essential for the
environmental cycles that regenerate
our planet.
Fields in Microbiology
◗ Medically related fields:
◗ Medical Microbiology
◗ Immunology
◗ Epidemiology
◗ Virology
◗ Molecular Pathogenesis

◗ General and Applied
Microbiology
◗ Environmental
Microbiology
◗ Molecular Microbiology
◗ Industrial Microbiology
◗ Biodiversity
◗ Bioremediation
◗ Food Microbiology
◗
What Are My Options?
◗ Microbiologists can specialize in a variety of areas. From the food to
the space industries, and everywhere in between, there are many
opportunities for microbiologists!


- Biochemist - Environmental Scientist

- Biotechnologist
- Bacteriologist
- Virologist
- Immunologist
- Mycologist
- Teacher
-

VIRUS FUNGI
What kind of work does an
industrial microbiologist do?
◗ The application of scientific and engineering principles to the
processing of materials by microorganisms (such as bacteria, fungi,
algae, protozoa and viruses) or plant and animal cells to create
useful products or processes.


◗ Responsible for the discovery, development, or implementation of

certain processes and the quality of resultant products:
- Antibiotics/Antimicrobials
- Vaccines
- Health care product
- Food & beverages produced my microbiology activity
-
What kind of work does an medical
microbiologist do?
◗ The study of microorganisms including bacteria, viruses, fungi and parasites
which are of medical importance and are capable of causing diseases in
human beings.
◗ Work with the diagnosis and management of infectious diseases in hospitals
and in the community and in preventing people at risk.



What kind of work does an food
microbiologist do?
◗ Food Industry ◗ Technical Sales and Marketing
 - Oversee and optimize the  - Participate in sales and marketing of
 conversion of raw materials into  ingredients, equipment and packaging
 finished products
 - Ensure product quality and safety  materials
 - Carry out new product  - Interface between suppliers and the
 development in a very competitive  food industry
 industry  - Provide scientific and technical
◗ Public Service  support
 - Develop / enforce legislation and
regulations related to food production
and safety
 - Inspect and evaluate food production
facilities
 - Improve post harvest technologies at
 home and abroad


◗
Where Does a Microbiologist Work
and With Whom?

Microbiologists work as integral members of

interdisciplinary teams in laboratories of hospitals, clinics,
universities, health care, research organization, industry
(food and drink, pharmaceutical, toiletry, and
biotechnology companies) and government.
The following lists provide a brief
sampling of the kind of jobs and
work environments
Sample Career Titles Sample Work Settings
Lab Technician Private Industry
Research Technician Hospitals
Clinical Microbiologist Private Industry
Microbiologist Education
Clinical Trials Manager Contract Research Organization
Lab Research Analyst Medical Research Laboratory

Ho
m
Plant Tissue
Culture
LABORATORY

MELAKA INSTITUTE OF
BIOTECHNOLOGY
 ACTIVITY


1. Research and Development of various

types of medicinal, ornamental and
forest plants species.


 RESEARCH OFFICER


1. Mrs. Azurin Hanim bt. Jamaluddin.

2. Mrs. Rabiatul Adawiyah bte Mohd. Yusof.
3.

 INTRODUCTION
 What is plant Tissue Culture???


 Plant tissue culture is a

practice used to propagate
plants under sterile
conditions, often to produce
clones of a plant
 INTRODUCTION
Plant tissue culture relies on the fact
: Any plant cells have the ability to
regenerate a whole plant(totipotency).

Parts of plants can be used to generate
a new plant are such as :
Single cells
Protoplasts (plant cells without
cell walls )
Pieces of leaves.
Roots
Stem

Explants was given with the required
nutrients and plant hormones included
 OBJECTIVE
 To produce/propagate :
 The exact copies of plants which have
desirable traits.
 Quickly mature plants.
 Multiples of plants in the absence of
seeds or necessary pollinators to
produce seeds.
 Whole plants from plant cells that have
been genetically modified.
 Plants with greatly reduced chances of
transmitting diseases, pests, and
pathogens.
 Plants which have very low chances of
germinating and growing, i.e.: orchids
 METHODS IN TISSUE
CULTURE
1.Media Preparations
2.Explants Preparations
Shoot induction
3.Culturing the explants
4. Callus induction

Shoot multiplication
5.Subculturing the plantlets
6. Root induction

7.Transplanting (Acclimatization)
 MEDIA PREPARATIONS

Addition of
Nutrients Measure pH
Weighing Sucrose

Addition of gelling
Autoclave Media cooking agents
E X P LA N T S ( se le ctio n a n d
ste riliza tio n )
C u ltu rin g th e exp la n ts
S u b cu ltu rin g th e
p la n tle ts
 Tra n sp la n tin g
( acclimatization )
Tra n sfe r to n u rse ry
( hardening )

Misai Kucing

Orchids

Chilli

Chilli

Rose Kacip
Orchids
Fatimah Sambung Nyawa
 O u r p ro d u cts

Sundal Malam

Misai Kucing Limau Kasturi

Misai Kucing

Orchids

Chilli

Hom
Nilam e
Sanserviera Trifasciata Teak
 FOOD
&
CHEMICAL
TESTING
LABORATORY

MELAKA INSTITUTE OF BIOTECHNOLOGY

‘Bridging Research with Industry’



 Offers various types of food testing:



i. Protein vii. Energy



ii.Total Fat viii. Benzoic Acid



iii.Moisture ix. Sorbic Acid



iv.Ash x. Saturated Fat

v.
vi.Carbohydrate xi. Cholestrol
vii.
viii.Vitamin A,C,D and E
Chemical Testing

i. Steroids
ii.
iii. Heavy Metals
iv.
v. Minerals
vi.
vii. Essential Oil
 Water Testing


i. Biochemical Oxygen Demand (BOD)

ii. Chemical Oxygen Demand (COD)
iii.pH
iv.Dissolve Oxygen (DO)
v. Total Soluble Solid (TSS)
vi.Total Dissolve Solid (TDS)
vii.Heavy Metals
viii.Minerals
Used in analytical and biochemistry to
separate, identify and quantify
compounds.
Hom
e
 Biology Fair
Kolej Matrikulasi Melaka

Toxicology Unit
Bridging research with industry

Melaka Institute of Biotechnology

Lot 7, MITC City, Hang Tuah Jaya, 75450 Ayer Keroh, Melaka.
06-231 3266
 Introduction

n Established in early 2007

n
n With 2 research officers
n
n Small laboratory with few
basic equipments –
Elisa plate reader for
cytotoxicity testing,
centrifuge, 160L fridge,
laminar flow cabinet for
mammalian cell sterile
working area,
fluorescence and
inverted microscope
and CO2 incubator for
culturing cell.
n
n In intention to expand the function of this lab, we have
proposed a project to MOSTI to develop a mutagenicity
testing facility in accordance to OECD GLP principle.
n
n On December 2007, MIB has been granted this project which
cost about RM4 million for 3 years.
n
n This facility for mutagenicity testing with GLP compliance is
estimate accomplished on 2010.
 Objectives
n Research for toxicology
n appointed as one of collabrators with
local/international research institute;
§ UiTM – toxicology profile for newly
synthesis stilbene


n Services - for local/international companies

manufacturing food products and supplements
n short term studies – offer services for in

vitro toxicology testing

n provide toxicology profile testing
 Provided Tests

¡ Mammalian cell line based;

i. In Vitro Cytotoxicity and Test (MTT Assay) and
mode of cell death determination
ii. In vitro Mammalian Chromosome Aberration Test
(OECD 473)
iii.In Vitro Mammalian Cell Gene Mutation Test
(OECD 476)
iv.In Vitro Sister Chromatid Exchange Assay in
Mammalian Cells (OECD 479)
v. Single Cell Gel Electrophoresis Assay (Comet
Assay)
vi.
¡ Bacteria cell based;
i. Bacterial Reverse Mutation Test (Ames Test)
(OECD 471)
ii. Umu Test
 What is
Toxicology?
n
n Toxicology (from the Greek words toxicos and logos) is
the study of the adverse effects of chemicals on living
organisms.


n It is an inter-disciplinary science that integrates the

principles and methods of many fields: chemistry,
biology, pharmacology, molecular biology, physiology
and medicine. It study the symptoms, mechanisms,
treatments and detection of poisoning, especially the
poisoning of people.
n
 Fields of
toxicology
n Risk assessment - attempts to determine,
probabilistically, outside the laboratory, the likelihood
of an adverse effect based on a particular exposure
scenario.
n
n

n Regulatory toxicology relies on risk assessment and

experimental data to determine the risk and benefits,
or the costs and benefits of exposure to certain
chemicals, to determine whether such chemical will
be allowed in the public sphere and to what extent its
use and exposure will be regulated.
n
n Clinical toxicology involves the application of
toxicological principles within a diagnostic setting,
usually to determine whether a presenting adverse
effect or disease or injury is due to some type of
chemical exposure.
n
n Forensic toxicology focuses on medical-legal aspects of
chemical exposure and toxic injury. Evaluation of
analytical chemistry techniques and measurements,
and testimony of conclusions and opinions in courts
of law encompass the activity of this specialized field
of toxicology.
n
n Aquatic toxicology is the study of the effects of
manufactured chemicals and other anthropogenic
and natural materials and activities on aquatic
organisms at various levels of organization, from
subcellular through individual organisms to
communities and ecosystems.
n
n Chemical toxicology is the study of the adverse
effects of chemicals on living organisms.
n
n Ecotoxicology is the integration of toxicology and
ecology or, as Chapman (2002) suggested,
“ecology in the presence of toxicants”. It aims
to quantify the effects of stressors upon natural
populations, communities, or ecosystems.
n
n Environmental toxicology and its sister-subject
ecotoxicology refers to the scientific study of
the potential effects upon organisms of
anthropogenic compounds released into the
natural environment.
n
 Why consider a
career in Toxicology?
n Challenges
n Wise use of chemicals is an essential component of
the high standard of living we enjoy. The
challenge to toxicologists is to ensure that we are
not endangering our health or the environment
with the products and by-products of modern and
comfortable living.


n As a career, toxicology provides the excitement of

science and research while also contributing to
the well-being of current and future generations.


n Opportunities
n With the increase in our health consciousness,
as well as concern for our environment, a
wide and growing variety of career
opportunities exist in toxicology.
n
n Attractive Salaries and Professional
Advancement
n The demand for well-trained toxicologists
continues to increase. Increasing
specialization in the science of toxicology now
provides the toxicologist with a competitive
advantage over chemists, engineers,
biologists or other scientists without
specialized training in toxicology.
 What do Toxicologists do?

¡ Research, Product Safety Evaluation,

Teaching, Public Service,
Regulatory Affairs and Consulting


¡ Where can I find more information?

l SOT – Society of Toxicology:
http://www.toxicology.org Hom
l SETAC – Society of Environmental
e
Toxicology and Chemistry:
http://www.setac.org/
 ‘Melaka Education Fair 2009’
Melaka
Institute of
Biotechnology

Thank you