1. Fundamentals of Biotechnology
Lecture # 3 Introduction & History of Biotechnology Recombinant Technology
By: Haji Akbar
M Phil

2. Biotechnology is most briefly
“the art of utilizing living organisms and their products for the production of food, drink, medicine or for other benefits to the human race, or other animal species”.

3. Humans have been making use of biotechnology since their life on earth.
e.g. Discovered farming,
Animal breeding
Cross-pollination of plants and cross-breeding of animals (used to enhance product quality and/or meet specific requirements or standards).

4. Most significant, life-altering discoveries.
The discovery of microorganisms and the subsequent burst of knowledge related to the causes of infectious diseases, antibiotics and immunizations.

5. Current scientific methods are more specific than historical techniques,
as scientists now directly alter genetic material using techniques known as recombinant DNA technology.

6. Subjects Involved With Biotechnology
Multidisciplinary- involving a number of disciplines that are coordinated for a desired outcome
Science
Life sciences
Physical sciences
Social sciences
Mathematics
Applied sciences
Computer applications
Engineering
Agriculture

7. Stages of Biotechnology Development
Ancient biotechnology- early history as related to food and shelter; Includes domestication
Classical biotechnology- built on ancient biotechnology; Fermentation promoted food production, and medicine
Modern biotechnology- manipulates genetic information in organism; Genetic engineering

8. Benefits of Biotechnology
Medicine
Human
Veterinary
Biopharming
Environment
Agriculture
Food products
Industry and manufacturing etc.

9. The legends in Biotechnology
Anton van Leeuwenhoek
(father of Microbiology)
Discovered cells

10. Gregor Johan Mendel
Discovered Genetics
(father of Genetics)

11. Walter Sutton
Discovered Chromosomes

12. Thomas Hunt Morgan
Discovered how genes are transmitted through chromosomes

13. Sir Alexander Fleming
Discovered penicillin

14. Rosalind Elsie Franklin
Research led to the discovery of the double helix structure of DNA

15. James Watson and Francis Crick
Discovered DNA

16. Ernst Ruska
Invented the electron microscope (in 1933).

17. Mary-Claire King
Mapped human genes for research of cancer treatments

18. Ian Wilmut
Created the first true clone, the Dorset ewe Dolly

19. Recombinant DNA Technology
Recombinant DNA (rDNA) technology is a field of molecular biology in which scientists "edit" DNA to form new synthetic molecules, which are often referred to as "chimeras".
Recombinant DNA technology works by taking DNA from two different sources and combining that DNA into a single molecule.
Recombinant DNA is a type of DNA that is artificially created by inserting a strand or more of DNA into a different set of DNA. often referred to as rDNA for short.

20. Recombinant DNA technology
Recombinant DNA technology is one of the recent advances in biotechnology, which was developed by two scientists named Boyer and Cohen in 1973.

21. Basic principle of recombinant DNA technology
The DNA is inserted into another DNA molecule called ‘vector’
The recombinant vector is then introduced into a host cell where it replicates itself

22. Steps involved recombinant DNA technology
1. DNA molecules are digested with enzymes called restriction endonucleases which reduces the size of the fragments  Renders them more manageable for cloning purposes
Can be called Restriction digestion

23. 2. These products of digestion are inserted into a DNA molecule called a vector 
Enables desired fragment to be replicated in cell culture to very high levels in a given cell (copy #)

24. 3. Introduction of recombinant DNA molecule into an appropriate host cell
Transformation or transfection
Each cell receiving rDNA = CLONE
May have thousands of copies of rDNA molecules/cell after DNA replication
As host cell divides, rDNA partitioned into daughter cells

25. 4. Population of cells of a given clone is expanded, and therefore so is the rDNA.
Amplification
DNA can be extracted, purified and used for molecular analyses
Investigate organization of genes
Structure/function
Activation
Processing
Gene product encoded by that rDNA can be characterized or modified through mutational experiments

26. Applications of Recombinant DNA Technology
Large-scale production of human proteins by genetically engineered bacteria.
Such as : insulin, Growth hormone, Interferons and
Blood clotting factors (VIII & IX)

27. Production of Human Insulin
1) Obtaining the human insulin gene
Human insulin gene can be obtained by making a complementary DNA (cDNA) copy of the messenger RNA (mRNA) for human insulin.

28. 2)Joining the human insulin gene into a plasmid vector
The bacterial plasmids and the cDNA are mixed together. The human insulin gene (cDNA) is inserted into the plasmid through complementary base pairing at sticky ends.

29. 3)Introducing the recombinant DNA plasmids into bacteria
The bacteria E.coli is used as the host cell. If E. coli and the recombinant plasmids are mixed together in a test-tube.

30. 4)Selecting the bacteria which have taken up the correct piece of DNA
The bacteria are spread onto nutrient agar. The agar also contains substances such as an antibiotic which allows growth of only the transformed bacteria. (selective media)

31. 2. Vaccine Development 3. recombinant factor 8 and recombinant factor 9 made by genetic engineering are now available for hemophilia A and B. 4. Gene therapy for genetic diseases