Lecture # 3Introduction & History of

BiotechnologyRecombinant Technology

Fundamentals of Biotechnology

By: Haji Akbar

M Phil

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”.

• 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).

• 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.

• Current scientific methods are more specific than historical techniques,

as scientists now directly alter genetic material using techniques known as recombinant DNA technology.

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

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

Benefits of Biotechnology

• Medicine– Human– Veterinary– Biopharming

• Environment• Agriculture• Food products• Industry and manufacturing etc.

The legends in Biotechnology

• Anton van Leeuwenhoek

(father of Microbiology)

• Discovered cells

• Gregor Johan Mendel• Discovered Genetics(father of Genetics)

• Walter Sutton

• Discovered Chromosomes

• Thomas Hunt Morgan• Discovered how genes

are transmitted through chromosomes

• Sir Alexander Fleming• Discovered penicillin

• Rosalind Elsie Franklin• Research led to the

discovery of the double helix structure of DNA

• James Watson and Francis Crick

• Discovered DNA

• Ernst Ruska• Invented the electron

microscope (in 1933).

• Mary-Claire King• Mapped human genes

for research of cancer treatments

• Ian Wilmut• Created the first true

clone, the Dorset ewe Dolly

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.

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.

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

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

• 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 #)

• 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

• 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

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)

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.

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.

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.

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)

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