Recombinant DNA Technology Pre-AP Biology 10/5/2015

Bellwork Explain what genetic engineering means to you. 10/5/2015

Key Terminology Recombinant Chimera Plasmid Cloning Vector Ligate / DNA Ligase Transformation Transgenic

Objectives Define recombinant DNA Explain how bacteria are genetically engineered Create a paper model of a recombinant plasmid and identify the key components Describe examples of Biotechnology in plants and animals 10/5/2015

Overview Recombinant DNA Technology creates new DNA molecules  made by piecing together DNA from different organisms  This recombinant DNA (rDNA) is also known as a chimera Goat boy

Examples of Recombinant DNA Technology rhInsulin  marketed by Eli, Lilly and Company Recombinant human growth hormone  Nutropin, marketed by Genentech, Inc Recombinant “Activase”  dissolves blockages in blood vessels  marketed by Genentech, Inc

Significance of rDNA These proteins normally are produced in very small amounts in animals Making these proteins in large amounts in other organisms, like bacteria  Can be supplemented in individuals that are deficient in a protein, like Insulin  Can be made cheaper and safer than other methods

Making a Recombinant DNA Molecule Step 1: Isolation of human DNA encoding Insulin:  Isolate Human DNA  Cut DNA into fragments with a restriction enzyme  Identify the DNA fragment that encodes for the protein Insulin

Making a Recombinant DNA Molecule Step 2: Place DNA molecule into a cloning vector  Cloning vector is commonly a plasmid small circular piece of DNA found in bacteria separate from its chromosomal DNA A plasmid is called a vector  when used to carry the desired DNA into the organism

Making a Recombinant DNA Molecule (Step 2 continued)  Fragments of DNA are put into the plasmid  Cut plasmid with same restriction enzyme used to make DNA fragments  Ligate fragments into the plasmid with DNA ligase DNA ligase “glues” the DNA fragments together

Think-Pair-Share Why is the plasmid with the human insulin DNA considered a recombinant DNA molecule or chimera? PROPERTY OF PIMA COUNTY JTED,

Transforming Bacteria with Recombinant DNA Step 3: The recombinant DNA molecule is taken up by the Bacteria  Bacteria are said to be transformed When they take up a foreign gene in a plasmid e.g. when they take up the human insulin gene

Transformation Process by which bacterium takes up and expresses foreign DNA Happens in nature when bacteria die and release their DNA, and other bacteria take up the DNA 13

Think-Pair-Share IYOW define transformation of bacteria. PROPERTY OF PIMA COUNTY JTED,

Identify the Bacteria with the Recombinant DNA molecule Not all bacteria will be transformed Must eliminate bacteria that do not carry the recombinant plasmid Accomplished by growing bacteria on media containing an antibiotic  Antibiotic –chemicals that prevent the growth and survival of bacteria How does the antibiotic work?

Definition 10/5/2015 Property of CTE Joint Venture 16 Antibiotic - “against life” substance that kills bacteria or prevents them from replicating

Cloning Vector The plasmid that is used as vector is also given an antibiotic resistance gene The vector shown has the resistant gene to ampicillin

How Bacteria Acquire Antibiotic Resistance Property of CTE Joint Venture 18

Bacteria Take Up Cloning Vector Bacteria that take up cloning vector will also be ampicillin resistant  In other words, transformed bacteria will not be killed when grown on agar containing ampicillin  The bacteria without the cloning vector will be killed because they do not contain the ampicillin resistance gene

Think-Pair-Share You have transformed bacteria with the plasmid containing the human insulin gene. Only 10% of the bacteria take up the recombinant DNA molecule. Predict which petri plate will have more bacteria growth, the one + ampicillin or the one – ampicillin. Explain your reasoning. PROPERTY OF PIMA COUNTY JTED,

Uses of Biotechnology in Plants Make Transgenic Plants to  Delay ripening-ex. Flavor-Saver Tomatoes

Uses of Biotechnology in Plants Improve nutritional content  e.g. Golden Rice Herbicide resistance  Coffee

Uses of Biotechnology in Animals  Be a pharmaceutical factory produce a large amount of an otherwise rare biological substance for medical use  Hormones  Blood clotting factors  Vaccines

Q & A: Recombinant DNA 1) Why is it useful to create bacteria that contain a new gene? 2) What is the purpose of restriction enzymes in this process? 3) What is the purpose of DNA ligase? 4) Describe the 3 steps for making recombinant DNA. 5) Human Growth Factor is a protein. Based on what you’ve learned in this unit, what biochemical processes must take place before HGH will be produced by bacteria?