1. Ch. 20 Warm-Up Share 3 things you are grateful for.
Use your textbook (Ch. 20) to answer the following review questions.
What is recombinant DNA?
What are plasmids?
What are restriction enzymes (RE)?
When DNA is cut using an RE, describe the ends of the DNA fragments.

2. Warm-Up
A bacterial plasmid is 100 kb in length. The plasmid DNA was digested to completion with 2 restriction enzymes in 3 separate treatments: EcoRI, HaeIII, and EcoRI + HaeIII (double-digest). The fragments were separated by gel electrophoresis below.
Draw a circle to represent the plasmid. On the circle, construct a labeled diagram of the restriction map of the plasmid.

3. Warm-Up
Describe how a plasmid can be genetically modified to include a piece of foreign DNA that alters the phenotype of bacterial cells transformed with the modified plasmid.
How can a genetically modified organism provide a benefit for humans and at the same time pose a threat to a population or ecosystem?

4. biotechnology
Chapter 20

5. Chapter 20: Terms to Know Genetic engineering Biotechnology
Recombinant DNA
Gene cloning
Plasmid
Restriction enzymes
Sticky ends
DNA ligase
Cloning vector
Nucleic acid hybridization
PCR
Gel electrophoresis
RFLPs
Genomic library
cDNA library
DNA microarray assays
SNPs
Stem cells
Gene therapy
Transgenic animals
GMO (genetically modified organism)

6. What You Must Know: The terminology of biotechnology.
How plasmids are used in bacterial transformation to clone genes.
The key ideas that make PCR possible and applications of this technology.
How gel electrophoresis can be used to separate DNA fragments or protein molecules.
Information that can be determined from DNA gel results, such as fragment sizes and RFLP analysis.

7. Genetic Engineering: process of manipulating genes and genomes
Biotechnology: process of manipulating organisms or their components for the purpose of making useful products.

9. Recombinant DNA: DNA that has been artificially made, using DNA from different sources
eg. Human gene inserted into E.coli
Gene cloning: process by which scientists can product multiple copies of specific segments of DNA that they can then work with in the lab

10. Tools of Genetic Engineering
Restriction enzymes (restriction endonucleases): used to cut strands of DNA at specific locations (restriction sites)
Restriction Fragments: have at least 1 sticky end (single-stranded end)
DNA ligase: enzyme that joins DNA fragments
Cloning vector: carries the DNA sequence to be cloned (eg. bacterial plasmid)

11. Using a restriction enzyme (RE) and DNA ligase to make recombinant DNA

12. Gene Cloning Using bacteria to clone a human gene

13. Applications of Gene Cloning

14. Genetically Modified organisms (GMO)
Organisms altered through recombinant DNA technology
Insert foreign DNA into genome or combine DNA from different genomes

16. Watch: Are GMOs Good or Bad? Genetic Engineering & Our Food
Take notes!

17. “Pharm” animal: produce human protein secreted in milk for medical use

18. Techniques of Genetic Engineering

19. Techniques of Genetic Engineering
Transformation: bacteria takes up plasmid (w/gene of interest); used in gene cloning
PCR (Polymerase Chain Reaction): amplify (copy) piece of DNA without use of cells
Gel electrophoresis: used to separate DNA molecules on basis of size and charge using an electrical current (DNA  + pole)
DNA microarray assays: study many genes at same time

20. PCR (Polymerase Chain Reaction): amplify (copy) piece of DNA without use of cells

21. PCR Virtual lab PCR – Genetic Science Learning Center
Click through the virtual lab and take notes.
Focus Points:
What is PCR?
How is it used?
What are the “pros”/benefits of this technique?
In general, how does it work?
What materials are needed?

22. Gel Electrophoresis: used to separate DNA molecules on basis of size and charge using an electrical current (DNA (-)  (+) pole)

23. Gel Electrophoresis
DNA Fingerprint

24. DNA Fingerprinting

25. Video: Gel electrophoresis
Take Notes!
Video: Gel electrophoresis
Amoeba sisters

26. DNA Restriction Enzyme Simulation
A formal lab grade!

27. Microarray Assay: used to study gene expression of many different genes

28. DNA microarray that reveals expression levels of 2,400 human genes

29. DNA MICROARRAY Virtual lab
DNA MICROARRAY – Genetic Science Learning Center
Click through Ch. 1 & 2 of the virtual lab and take notes as needed.
For Ch. 3 (The Experiment) take notes on these Focus Points:
What is a DNA Microarray
How is it used?
What are the “pros”/benefits of this technique?
In general, how does it work?
What materials are needed?

30. Cloning Organisms
Nuclear transplantation: nucleus of egg is removed and replaced with nucleus of a body cell

31. Nuclear Transplantation

32. Problems with Reproductive Cloning
Cloned embryos exhibited various defects
DNA of fully differentiated cell have epigenetic changes
Human Cloning

33. Is it cloning or not?
Go to Genetic Science Learning Center and play the Quiz Game “Is it Cloning or Not”

34. Stem Cells
Stem cell: can reproduce itself indefinitely and produce other specialized cells
Zygote = totipotent (able to differentiate into any type of cell)
Embryonic stem cells = pluripotent (can become many cell types)
Adult stem cells = multipotent (can become a few cell types) or induced pluripotent, iPS (forced to be pluripotent)

35. Embryonic vs. Adult stem cells

36. Using stem cells for disease treatment

37. Explore stem cells Genetic Science Learning Center
Click through the links and videos under “Stem Cells”
Take notes on these Focus Points:
What different kinds of stem cells are there
Benefits and Drawbacks to using/obtaining different stem cells types
Ethical considerations for stem cell use
Uses of stem cells

38. Gene therapy using a retroviral vector

39. Explore GENE THERAPY Genetic Science Learning Center
Click through the links and videos under “Gene Therapy”
Take notes on these Focus Points:
What are vectors?
What different types of vectors are there? (visit the vector toolbox)
What types of disorders/diseases are good targets for gene therapy? (and why?)
Challenges of gene therapy
Approaches to gene therapy (3 ways (not names))

40. RFLPs (“rif-lips”) Restriction Fragment Length Polymorphism
Cut DNA with different restriction enzymes
Each person has different #s of DNA fragments created
Analyze DNA samples on a gel for disease diagnosis
Outdated method of DNA profiling (required a quarter-sized sample of blood)

41. RFLPs – Disease Diagnosis

42. STR Analysis STR = Short Tandem Repeats
Non-coding DNA has regions with sequences (2-5 base length) that are repeated
Each person has different # of repeats at different locations (loci)
Current method of DNA fingerprinting used – only need 20 cells for analysis

43. STR Analysis

44. STR Analysis

45. Applications of DNA Technology
Diagnosis of disease – identify alleles, viral DNA
Gene therapy – alter afflicted genes
Production of pharmaceuticals
Forensic applications – DNA profiling
Environmental cleanup – use microorganisms
Agricultural applications - GMOs