1. BIOTECHNOLOGY

2. Dolly and surrogate Mom
Biotechnology
6/24/2018
EXAMPLES
Dolly and surrogate Mom
Embryonic stem cells and gene therapy
Genetically modified rice.
G. Podgorksi, Biol. 1010

3. Biotechnology
6/24/2018
Biotechnology
Biotechnology, defined broadly, is the engineering of organisms for useful purposes.
Often, biotechnology involves the creation of hybrid genes and their introduction into organisms in which some or all of the gene is not normally present.
Fourteen month-old genetically engineered (transgenic) salmon (left) and standard salmon (right).
G. Podgorksi, Biol. 1010

4. Biotechnology We’ll examine: Reproductive & Therapeutic Cloning
6/24/2018
Biotechnology
We’ll examine:
Reproductive & Therapeutic Cloning
Genetically modified foods
Gene cloning for pharmaceutical production
DNA fingerprinting
The promise and perhaps perils of embryonic stem cells
Gene therapy
G. Podgorksi, Biol. 1010

5. Animal Cloning Dolly and her surrogate mother. Biotechnology 6/24/2018
G. Podgorksi, Biol. 1010

6. Why Clone Animals? To answer questions of basic biology
Biotechnology
Why Clone Animals?
6/24/2018
Five genetically identical cloned pigs.
To answer questions of basic biology
For pharmaceutical production.
For herd improvement.
To satisfy our desires (e.g. pet cloning).
G. Podgorksi, Biol. 1010

7. Cloning by Somatic Cell Nuclear Transfer (SCNT)
Biotechnology
6/24/2018
Cloning by Somatic Cell Nuclear Transfer (SCNT)
page 185
This is the first step in both therapeutic and reproductive cloning. Reproductive cloning is the cloning of whole organisms (usually for agricultural purposes). Therapeutic cloning is the cloning of stem cells to be used for therapeutic purposes.
ANIMATIONS
G. Podgorksi, Biol. 1010

8. Is Reproductive Cloning Ethical?
Biotechnology
6/24/2018
Is Reproductive Cloning Ethical?
The Next Step?
Highly unlikely.
Attempts at human cloning are viewed very unfavorably in the scientific community.
G. Podgorksi, Biol. 1010

9. Recombinant DNA DNA can be cut at specific sequences using restriction
Biotechnology
6/24/2018
Recombinant DNA
DNA can be cut at specific sequences using restriction
enzymes.
This creates DNA fragments useful for manipulating
and splicing genes, so that completely novel DNA
combinations can be created.
Most of these new DNA combinations would
never have occurred naturally.
These are mature and widely utilized biotechnologies.
G. Podgorksi, Biol. 1010

10. Biotechnology
6/24/2018
Restriction Enzymes are Enzymes That Cut DNA Only at Particular Sequences
The enzyme EcoRI cutting DNA at its recognition sequence
Restriction enzyme animation
Different restriction enzymes have different recognition sequences.
This makes it possible to create a wide variety of different gene fragments.
G. Podgorksi, Biol. 1010

11. DNAs Cut by a Restriction Enzyme Can be Joined Together in New Ways
Biotechnology
6/24/2018
DNAs Cut by a Restriction Enzyme Can be Joined Together in New Ways
These are recombinant DNA molecules.
Each colour is DNA from a different species.
If inserted into an organism, the resulting organism is referred to as a genetically modified organism (GMO) or a transgenic organism. (page 187)
G. Podgorksi, Biol. 1010

12. Plasmids are Used to Replicate a Recombinant DNA
Biotechnology
6/24/2018
Plasmids are Used to Replicate a Recombinant DNA
Plasmids are small circles of DNA found in bacterial cells.
Plasmids replicate independently of the bacterial chromosome.
Pieces of foreign DNA can be added within a plasmid to create a recombinant plasmid.
Replication often produces copies of a recombinant plasmid in each cell.
G. Podgorksi, Biol. 1010

13. Overview of GENE CLONING
Biotechnology
6/24/2018
Overview of GENE CLONING
PAGE 184
ANIMATION
Cloning animation
G. Podgorksi, Biol. 1010

14. Biotechnology
Harnessing the Power of Recombinant DNA Technology – Human Insulin Production by Bacteria
6/24/2018
G. Podgorksi, Biol. 1010

15. Human Insulin Production by Bacteria
Biotechnology
Human Insulin Production by Bacteria
6/24/2018
and cut with a restriction enzyme
6) join the plasmid and human fragment
G. Podgorksi, Biol. 1010

16. Human Insulin Production by Bacteria
Biotechnology
Human Insulin Production by Bacteria
6/24/2018
Mix the recombinant plasmid with bacteria.
Screening bacterial cells to learn which contain the human insulin gene is the hard part.
G. Podgorksi, Biol. 1010

17. Route to the Production of Human Insulin by Bacteria
Biotechnology
Route to the Production of Human Insulin by Bacteria
6/24/2018
One cell with the recombinant plasmid
A fermentor used to grow recombinant bacteria.
This is the step when gene cloning takes place.
The single recombinant plasmid replicates within a cell.
Then the single cell with many recombinant plasmids produces trillions of like cells with recombinant plasmid – and the human insulin gene.
G. Podgorksi, Biol. 1010

18. Route to the Production of Human Insulin by Bacteria
Biotechnology
Route to the Production of Human Insulin by Bacteria
6/24/2018
The final steps are to collect the bacteria, break open the cells, and purify the insulin protein expressed from the recombinant human insulin gene.
Prior to this technology, human diabetics injected themselves with purified pork insulin.
G. Podgorksi, Biol. 1010

19. Biotechnology
Pharming
6/24/2018
Pharming is the production of pharmaceuticals in animals engineered to contain a foreign, drug-producing gene (page 188).
Example: edible vaccines
G. Podgorksi, Biol. 1010

20. The Promise and Possible Perils of Stem Cells
Biotechnology
6/24/2018
The Promise and Possible Perils of Stem Cells
G. Podgorksi, Biol. 1010

21. Biotechnology
6/24/2018
Stem Cells
A stem cell is an undifferentiated, dividing cell that gives rise to a daughter cell like itself and a daughter cell that becomes a specialized cell type.
Pages 186 & 189
G. Podgorksi, Biol. 1010

22. Biotechnology
6/24/2018
Stem Cells are Found in the Adult, but the Most Promising Types of Stem Cells for Therapy are Embryonic Stem Cells
G. Podgorksi, Biol. 1010

23. Some Thorny Ethical Questions
Biotechnology
6/24/2018
Some Thorny Ethical Questions
Are these masses of cells a human?
Is it ethical to harvest embryonic stem cells from the “extra” embryos created during in vitro fertilization?
G. Podgorksi, Biol. 1010

24. Biotechnology
6/24/2018
Additional Potential Dilemmas – Therapeutic Cloning to Obtain Matched Embryonic Stem Cells
Cells from any source other than you or an identical twin present the problem of rejection.
If so, how can matched embryonic stem cells be obtained?
A cloned embryo of a person can be made, and embryonic stem cells harvested from these clones.
G. Podgorksi, Biol. 1010

25. Is there any ethical difference between therapeutic
Biotechnology
6/24/2018
Therapeutic Cloning
Is there any ethical difference between therapeutic
and reproductive cloning?
G. Podgorksi, Biol. 1010

26. DNA, the Law, and Many Other Applications
Biotechnology
DNA Fingerprinting
DNA, the Law, and Many Other Applications
6/24/2018
A DNA fingerprint used in a murder case.
The defendant stated that the blood on his clothing was his.
What are we looking at? How was it produced?
G. Podgorksi, Biol. 1010

27. DNA Fingerprinting Basics
Biotechnology
DNA Fingerprinting Basics
6/24/2018
Different individuals carry different alleles.
Most alleles useful for DNA fingerprinting differ on the basis of the number of repetitive DNA sequences they contain.
If DNA is cut with a restriction enzyme that recognizes sites on either side of the region that varies, DNA fragments of different sizes will be produced.
A DNA fingerprint is made by analyzing the sizes of DNA fragments produced from a number of different sites in the genome that vary in length.
The more common the length variation at a particular site and the greater the number the sites analyzed, the more informative the fingerprint.
G. Podgorksi, Biol. 1010

28. A Site With Three Alleles Useful for DNA Fingerprinting
Biotechnology
6/24/2018
A Site With Three Alleles Useful for DNA Fingerprinting
DNA fragments of different size will be produced by a restriction enzyme that cuts at the points shown by the arrows.
G. Podgorksi, Biol. 1010

29. Biotechnology
6/24/2018
The DNA Fragments Are Separated on the Basis of Size using GEL ELECTROPHORESIS
The pattern of DNA bands is compared between each sample loaded on the gel.
Gel electrophoresis animation
G. Podgorksi, Biol. 1010

30. Possible Patterns for a Single “Gene” With Three Alleles
Biotechnology
6/24/2018
Possible Patterns for a Single “Gene” With Three Alleles
In a standard DNA fingerprint for a legal case, about a dozen sites are analyzed, with each site having many possible alleles.
G. Podgorksi, Biol. 1010

31. (i.e. make sure the cell has enough materials)
Biotechnology
6/24/2018
Which parental DNA matches the child’s? How did you decide this?
Determine the percentage of the father’s DNA and mother’s DNA that matches the child’s. Explain this.
ells start preparing for synthesis phase by undergoing various metabolic processes
(i.e. make sure the cell has enough materials)
S Phase / Synthesis Phase
DNA is replicated / copied
G2 / Gap 2
Cells are preparing to undergo division
Division Stage
(a) Mitosis – division of cell’s nucleus
(b) Cell division – division of cytoplasm
G. Podgorksi, Biol. 1010

32. Genetically Modified Foods
Biotechnology
6/24/2018
Genetically Modified Foods
Golden Rice is Modified to be Provide a Dietary Source of Vitamin A (Page 187)
Worldwide, 7% of children suffer vitamin A deficiency, many of them living in regions in which rice is a staple of the diet.
Many of our crops in North America are genetically modified.
Should they be?
G. Podgorksi, Biol. 1010

33. Biotechnology
6/24/2018
Methods for Plant Genetic Engineering are Well-Developed and Similar to Those for Animals
G. Podgorksi, Biol. 1010

34. GMOs, Especially Outside the US, Are a Divisive Issue
Biotechnology
6/24/2018
GMOs, Especially Outside the US, Are a Divisive Issue
Europeans have legislated mandatory labeling of GM products, but not in North America
Current Concerns Focus on the Environmental & Health Effects of GM Crops
The jury’s still out on the magnitude of GM crop’s ecological and health impacts, but the question is debated seriously.
G. Podgorksi, Biol. 1010

35. GENE THERAPY using a virus (Trojan Horse) page 226
Viruses are very specific for their host cells
Insertion of new gene into host genome is random