BIOTECHNOLOGY

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

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

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

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

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

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 http://www.dnatube.com/video/563/Somatic-Cell-Nuclear-Transfer-Animation http://learn.genetics.utah.edu/content/tech/cloning/clickandclone/ G. Podgorksi, Biol. 1010

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

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

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

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

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 50-100 copies of a recombinant plasmid in each cell. G. Podgorksi, Biol. 1010

Overview of GENE CLONING Biotechnology 6/24/2018 Overview of GENE CLONING PAGE 184 ANIMATION http://highered.mcgraw-hill.com/sites/0072556781/student_view0/chapter14/animation_quiz_1.html Cloning animation G. Podgorksi, Biol. 1010

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

(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

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

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

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

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 http://www.dnatube.com/video/4294/Gene-Therapy--cure-for-genetic-disorder