Chapter 3 Lesson 1

3.1.1 Differentiate, using examples, between genetic engineering and biotechnology
3.1.2 Summarize the roles of DNA tools (Include: restriction enzymes and gel electrophoresis)
3.1.3 Interpret data collected from the process of gel electrophoresis
3.1.4 Sequence, using diagrams and models, how recombinant DNA is made and manipulated
3.1.5 Explain the role of plasmids and DNA ligase in recombinant DNA technology
3.1.6 Describe in steps, using a diagram, the process of gene cloning
3.1.7 Describe in steps, using a diagram, the process of DNA sequencing
3.1.8 Detail, using a diagram, the steps of PCR
3.1.9 Analyze the application of biotechnology (transgenic organisms) in plants, animals, and bacteria

Genetic Engineering
- Definition: Technology involving the manipulation of the DNA of one organism to insert exogenous DNA (from another organism).
- Example: Introducing a gene from one species into another.
Biotechnology
- Definition: The use of genetic engineering to find solutions to problems. It allows the production of organisms containing individual genes from other organisms, hence creating transgenic organisms.
- Applications:
- Studying gene expression.
- Investigating cellular processes.
- Studying disease development.
- Selecting traits for human benefit.

Biotechnology: Involves using living organisms for human benefit.
Genetic engineering: A branch of biological science that alters genetic material directly.

Bacteria Applications:
- Produce substances like insulin, growth hormones, and anticoagulants.
Human Applications:
- Examples of substances produced:
- Lysosome
- Lactoferrin
Plant Applications:
- Engineered for greater resistance to pests and diseases.
- Examples:
- Sweet-potato resistant to viruses threatening African crops.
- Rice enriched with iron and vitamins to combat malnutrition.
- Plants engineered for tolerance to extreme weather.

Definition: Enzymes that recognize specific DNA sequences, binding and cleaving the DNA within those sequences.
Function:
- Create DNA fragments with sticky or blunt ends for further manipulation.
- Also known as endonucleases, powerful for isolating specific genes or genomic regions, creating fragments of variable sizes unique to each individual.
Sticky Ends vs. Blunt Ends:
- Sticky Ends: Created by some restriction enzymes like EcoRI, which cuts at specific sequences (GAATTC), yielding fragments with unpaired single-stranded DNA.
- Blunt Ends: Produced when restriction enzymes cut straight across both strands, lacking single-stranded regions, allowing them to join any other blunt-ended fragments.

Definition: Technique for separating DNA fragments based on size using an electric current.
Process:
- DNA is loaded into a gel and subjected to an electric current, moving towards the positive pole due to its negative charge.
- Smaller fragments migrate faster than larger ones, creating a banding pattern indicative of the fragment sizes.
- Bands can be stained for visibility and analyzed for comparisons or further studies.

Criminal Case Interpretation: Forensic analysis using band patterns from gel electrophoresis can help draw conclusions in criminal investigations. This might include comparing DNA from different sources (e.g., defendant vs. victim).

Definition: Combining DNA fragments from different sources to create recombinant DNA molecules.
Process:
1. DNA Cutting: DNA is cut using restriction enzymes.
2. Vector Cutting: The vector (e.g., plasmid) is also cut with the same enzyme.
3. Combination: Joining DNA fragments with a vector using DNA ligase.

Definition: Small, circular double-stranded DNA molecules naturally found in bacteria and yeast, suitable for carrying DNA into target cells.
Characteristics: Typically contain:
- Origin of replication (ori)
- Genetic markers (like antibiotic resistance genes)
- Restriction enzyme cutting sites (e.g., EcoRI).

Process Description:
1. Recombinant plasmids are introduced into bacterial cells through a process called transformation.
2. Transformation methods include electric pulsation or heat shock to create openings in the bacterial membrane, allowing plasmid entry.
3. Selection: An antibiotic resistance gene (e.g., ampicillin) is included in the plasmid for selecting transformed bacteria; only those that take up the plasmid survive.

Purpose: To determine the nucleotide sequence of DNA, predict gene function and identify mutations.
Process:
1. Mix unknown DNA fragment with DNA polymerase and nucleotides (A, G, C, T).
2. Use fluorescently tagged nucleotides that stop the reaction upon incorporation into the DNA strand, producing varied length fragments.
3. Separate tagged strands using gel electrophoresis and analyze using sequencing machines to determine the original DNA sequence.

Definition: A technique used to amplify specific DNA fragments, making millions of copies for analysis.
Steps of PCR:
1. Denaturation: Heating separates DNA strands.
2. Annealing: Cooling allows primers to attach to DNA strands.
3. Extension: DNA polymerase binds and adds nucleotides, repeating the cycle 20-40 times to yield millions of copies.

Definition: Organisms that contain DNA from another organism due to genetic engineering.

Medical:
- Transgenic goats producing antithrombin III (blood clotting prevention).
- Disease-resistant animals (turkeys and chickens).
- Fast-growing fish used for agricultural purposes.
Agricultural:
- Insect and herbicide-resistant plants.
- Virus-resistant crops and plants able to endure extreme weather.
Bacterial Applications:
- Production of therapeutic proteins like insulin.
- Bacteria engineered to protect crops from frost and clean up environmental pollutants.

Selective Breeding: Utilizes naturally occurring gene variants within a species through traditional breeding methods.
Genetic Engineering: Direct laboratory manipulation of an organism's genome to create genetic changes that may not occur naturally.