WEEK 1 – GENETIC ENGINEERING AND APPLICATION OF RECOMBINANT DNA MOST ESSENTIAL

GENETIC ENGINEERING
• Genetic engineering is the direct modification of an organism’s genome, which is the list of specific traits (genes)
stored in the DNA.

A. Classical Breeding
– Classical breeding, also known as traditional breeding or conventional breeding, is a method of improving the
genetic makeup of an organism by selectively mating individuals with desirable traits.
• selective breeding: Selective breeding involves choosing specific individuals with desirable traits to be
parents of the next generation. It relies on human intervention to control the mating of organisms.
• hybridization: Hybridization involves the controlled mating of two different but closely related species
or varieties to produce offspring with a combination of traits from both parents.
• inbreeding: Inbreeding is the mating of closely related individuals within the same breeding
population.

Steps in Classical Breeding

Step 1: Identify desirable traits. The first step in the classical breeding process is to identify the traits that are desirable
for the organism. In the case of corn, breeders were looking for traits such as increased yield, disease resistance, and
improved drought tolerance.

Step 2: Selection of parents. Once the desirable traits have been identified, the next step is to select parents for
breeding. The parents should be individuals that possess the desired traits to a high degree. In the case of corn,
breeders selected corn plants that were high-yielding and resistant to diseases and pests.

Step 3: Crossbreeding. The parents are then crossed, meaning that they are mated with each other. The offspring of
this cross will have a combination of the genes from both parents. In the case of corn, breeders crossed high-yielding
corn plants with disease-resistant corn plants.

Step 4: Selection of offspring. The offspring are then evaluated for the desired traits. The individuals that exhibit the
most desirable traits are then selected for further breeding. In the case of corn, breeders selected corn plants that were
high-yielding, disease-resistant, and drought-tolerant.

Step 5: Repeat steps 2-4. The classical breeding, the modification or alteration of the traits of an organism is indirectly.

B. Recombinant DNA Technology

Recombinant DNA technology, also known as genetic engineering, involves the manipulation of DNA to create new
combinations of genetic material that would not naturally occur.

Step 1: Isolation of DNA

 DNA is extracted from the organism of interest, using methods that can include breaking open cells,
separating cellular components, and purifying the DNA.

Step 2: Cutting and Splicing DNA

 Enzymes called restriction enzymes are used to cut DNA at specific recognition sites. The cut DNA fragments can
then be combined with DNA from another source.

Step 3: Inserting Foreign DNA into Plasmid

 The desired gene or DNA fragment is then inserted into a vector, which is a small, self-replicating DNA
molecule.
Step 4: Joining DNA Fragments

 The DNA fragment and the vector are then joined together using an enzyme called DNA ligase.

Step 5: Transformation
 It is a process of introducing the recombinant DNA molecule into a host cell, typically a bacterium.

Step 6: Selection and Screening of Transformed Cells

 The transformed cells are then selected based on a marker gene that is present on the vector. This marker
gene allows for the identification of cells that have successfully taken up the recombinant DNA.
Step 7: Amplification of recombinant DNA
 The recombinant DNA is then amplified by growing the transformed host cells in culture. This produces
multiple copies of the recombinant DNA molecule.

Step 8: Purification of Recombinant DNA

 The recombinant DNA is then purified from the host cells using various purification techniques.

Step 9: Analysis of recombinant DNA

 The purified recombinant DNA is then analyzed to ensure its integrity and to confirm the presence of the
desired gene or DNA fragment.

Step 10: Expression of Recombinant DNA

 The recombinant DNA can be introduced into a suitable expression host for the production of the desired
protein or product.

II. APPLICATION OF RECOMBINANT DNA

A. Agriculture
• Enhanced Nutritional Content
– Biofortification: Genetic engineering is employed to enhance the nutritional content of crops.
• Improved Crop Yield
– Recombinant DNA technology has enabled the development of genetically modified (GM) crops that
produce higher yields.
• Genetically Engineered Traits
– Herbicide Resistance
• Certain crops have been engineered to be resistant to specific herbicides, allowing for
effective weed control without harming the crop.
– Pest and Disease Resistance:
• GM crops can be engineered to resist pests and diseases, reducing the need for pesticides and
herbicides.
– Virus Resistance
• Plants can be modified to resist viral infections, reducing the need for chemical pesticides and
increasing crop yield and quality.
– Tolerance to Environmental Stresses
• GM crops can be developed to withstand environmental stresses such as drought, salinity, and
extreme temperatures.

B. Medicine
• Biopharmaceuticals
– Insulin Production
• Recombinant DNA technology has enabled the production of human insulin in bacteria, yeast, or
mammalian cells.
• This has replaced the traditional extraction of insulin from animal pancreases, ensuring a more
abundant and safer supply.
• Vaccine Development
– Subunit Vaccines
• Recombinant technology is used to produce subunit vaccines, which contain only the specific
antigens needed to stimulate an immune response.
• This allows for safer and more precise vaccine development.

• Gene Therapy
– Treatment of Genetic Disorders
• Recombinant DNA is employed in gene therapy to treat genetic disorders by introducing
functional genes into a patient's cells, correcting or compensating for the genetic defect.

• Production of Therapeutic Proteins

– Monoclonal Antibodies
• Monoclonal antibodies, vital for various medical treatments, are produced using recombinant DNA
technology.
• These antibodies can be designed to target specific diseases, such as cancer.
C. Industry
• Enzyme Production
– Industrial Enzymes
• Recombinant DNA technology is used to produce enzymes with specific industrial
applications, such as the production of detergents, textiles, and biofuels.
• Industry
• Bioremediation
– Environmental Cleanup
• Engineered microorganisms can be used for bioremediation, breaking down pollutants in soil and
water.
• This technology has applications in cleaning up oil spills, heavy metal contamination, and other
environmental hazards.
• Pharmaceutical Production
– Recombinant Protein Drugs
• The pharmaceutical industry utilizes recombinant DNA technology to produce therapeutic
proteins and drugs, including hormones and clotting factors.
• Biosensors
– Diagnostic Tools
• Recombinant DNA technology is employed in the development of biosensors for the detection of
various substances, aiding in medical diagnostics and environmental monitoring.
• Recombinant DNA technology is used to produce bioplastics from renewable resources, such as
plant-based materials. Bioplastics are biodegradable and offer an eco-friendly alternative to
traditional plastics.
• Production of Bioplastic
• Recombinant DNA technology is used to produce bioplastics from renewable resources, such as
plant-based materials.
• Bioplastics are biodegradable and offer an eco-friendly alternative to traditional plastics.