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Karnival Pendidikan Melaka 2009
Karnival Pendidikan Melaka 2009
Biology MoleculeToxicologyChemistry & Food TechnologyPlant Tissue CultureMicrobiology
Biology MoleculeToxicologyChemistry & Food TechnologyPlant Tissue CultureMicrobiology
Applications of Molecular Biology Techniques in Biotechnology
Industry
Amelia Kassim
Nurhuda Jalil
Nurul Arneida Husin
‘Education Fair 2009’ Melaka Institute of Biotechnology
Molecular biology is the study of biology at a molecular level.
INTRODUCTION
One of the most basic techniques of molecular biology to study protein function is expression cloning
Melaka Institute of Biotechnology
Plant Tissue Culture
Microbiology
Disciplines relationMarket intelligence
BioinformaticProteomic
What is DNA cloning?
When DNA is extracted from an organism, all its genes are obtained
In gene (DNA) cloning a particular gene is copied (cloned)
• 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
What is Gene cloning?
How is DNA cloned?
Cell-based DNA cloning Cell-free DNA cloning (PCR)
Clone in dividing cells
First cycle
Primer oligonucleotide
DNA replication
Denaturation, annealing
DNA sequence to be amplified
Second cycles
20 – 30 cycles
Amplified DNA sequences
Clone in PCR
Some Molecular Biology Techniques1. DNA / RNA extraction2. pDNA purification3. Polymerase chain reaction (PCR)4. Expression cloning5. Gel electrophoresis6. Gel documentation, and image analysis7. Southern blotting8. Northern blotting9. Western blotting and immunochemistry10. cDNA / genomic DNA library construction, and its screening 11. Southern, and Northern Hybridization techniques12. 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 transformation19. Analysis of the transgenic plants 20. Microscopy21. Cultivation of bacteria / Microbiology, 22. preparation of glycerol stocks23. Generation of ESTs24. Analysis of DNA, cDNA / ESTs sequences using online and offline bioinformatics tools 25. Promoter isolation and characterization ++++++++++++++++++++++++++++++
When you are working in
Molecular Biology Lab
Melaka Institute of Biotechnology
Gel-Documentation
RT-PCR
Spectrophotometer
Melaka Institute of Biotechnology
Gel Electrophoresis
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 ++
Metabolic Engineering
Power of Metabolic Engineering
Genetically modified
DNA FINGERPRINTIN
G TECHNOLOGY
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.
A-T
G-C
T-A
C-G
C-G
G-C
A-T
BASE PAIRING
A- ADENINE
T- THYMINE
G- GUANINE
C- CYTOSINE
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
Raw materials
Isolation of Genomic DNA
Loading PCR product
Selection of Primer/s
Adding PCR Component and tested samples
Run PCR
PCR product
Run Gel Electrophoresis
Unique DNA Fingerprint profiles
Data analysis
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.
• 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
Roles of Molecular Markers in Herbal Drug Technology?
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.
Microbiology World and You
MICROBIOLOGY UNIT
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
FUNGIVIRUS
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 - Oversee and optimize the conversion of raw materials into finished products - Ensure product quality and safety - Carry out new product development in a very competitive industry Public Service - 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
Technical Sales and Marketing - Participate in sales and marketing of ingredients, equipment and packaging materials - Interface between suppliers and the food industry - Provide scientific and technical support
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
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
2. Mrs. Azurin Hanim bt. Jamaluddin.3. Mrs. Rabiatul Adawiyah bte Mohd.
Yusof.
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
INTRODUCTIONPlant 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 on culture media in order to propagate.
OBJECTIVETo 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 CULTURE1.Media Preparations 2.Explants Preparations3.Culturing the explants
4.Subculturing the plantlets
5.Transplanting (Acclimatization)
Shoot multiplication
Root induction
Shoot induction
Callus induction
MEDIA PREPARATIONS
Addition of Nutrients
Weighing Sucrose Measure pH
Addition of gelling agentsMedia cooking Autoclave
PREPARATIONS OF EXPLANTS (selection and sterilization)
Culturing the explants
Subculturing the plantlets
Transplanting (acclimatization)
Transfer to nursery (hardening)
Chilli
Sambung Nyawa
Orchids
Misai Kucing
Orchids
Chilli
Kacip FatimahRose
Our products
Misai Kucing
Sanserviera Trifasciata Teak
Nilam
Limau Kasturi
Sundal Malam
Chilli
Orchids
Misai Kucing
FOOD &
CHEMICAL TESTING
LABORATORY
MELAKA INSTITUTE OF BIOTECHNOLOGY
‘Bridging Research with Industry’
ACTIVITY1. Research and Development of Natural Plant and
Herb2. Food and Chemical Analysis
Research Officer3. Ms.Ong Pei Thing, AMIC4. Cik Sabrina Hj Md Kassim5. En. Musalman Rambeli
Natural Plants and Herbs
1. Screening of various local plants and herbs
for antioxidant properties.
2. Production of antioxidant-rich extract from plants and herbs.
3. Development of nutraceutical and cosmeceutical products from potential plants.
4. New formulation of Biopesticide on Vegetable Farming.
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. Carbohydrate xi. Cholestrol
vi. Vitamin A,C,D and E
Chemical Testing
i. Steroids
ii. Heavy Metals
iii. Minerals
iv. 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
Gas Chromatograph / Mass Spectrometer (GC/MS)
i. Used in detecting and analyzing the chemical compound typically from organic compound.
Used in analytical and biochemistry to separate, identify and quantify compounds.
a measurement of the dissolved sugar-to-water mass ratio of a liquid.
Efficient method to remove solvent from sample by evaporating usually used with plant and herbs methanolic extracts.
Toxicology Unit
Bridging research with industry
Melaka Institute of BiotechnologyLot 7, MITC City, Hang Tuah Jaya, 75450 Ayer Keroh, Melaka.06-231 3266
Biology Fair
Kolej Matrikulasi Melaka
Established in early 2007
With 2 research officers
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.
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.
On December 2007, MIB has been granted this project which cost about RM4 million for 3 years.
This facility for mutagenicity testing with GLP compliance is estimate accomplished on 2010.
Research for toxicology appointed as one of collabrators with
local/international research institute; UiTM – toxicology profile for newly synthesis
stilbene
Services - for local/international companies manufacturing food products and supplements
short term studies – offer services for in vitro toxicology testing
provide toxicology profile testing
Mammalian cell line based;i. In Vitro Cytotoxicity and Test (MTT Assay) and mode
of cell death determinationii. 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)
Bacteria cell based;i. Bacterial Reverse Mutation Test (Ames Test) (OECD
471)ii. Umu Test
Toxicology (from the Greek words toxicos and logos) is the study of the adverse effects of chemicals on living organisms.
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.
Risk assessment - attempts to determine, probabilistically, outside the laboratory, the likelihood of an adverse effect based on a particular exposure scenario.
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.
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.
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.
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.
Chemical toxicology is the study of the adverse effects of chemicals on living organisms.
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.
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.
Challenges 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.
As a career, toxicology provides the excitement of science and research while also contributing to the well-being of current and future generations.
Opportunities 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.
Attractive Salaries and Professional Advancement 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.
Research, Product Safety Evaluation, Teaching, Public Service, Regulatory Affairs and Consulting
Where can I find more information? SOT – Society of Toxicology:
http://www.toxicology.org SETAC – Society of Environmental Toxicology
and Chemistry: http://www.setac.org/
Thank you
‘Melaka Education Fair 2009’
Melaka Institute of Biotechnology