recombinant dna technology pre-ap biology 10/5/2015
TRANSCRIPT
Recombinant Recombinant DNA DNA
TechnologyTechnology
Pre-AP Biology
04/19/23
BellworkBellwork Explain what genetic
engineering means to you.
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Key Terminology Recombinant Chimera Plasmid Cloning Vector Ligate / DNA Ligase Transformation Transgenic
ObjectivesObjectives
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
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Overview
Recombinant DNA Technology creates new DNA molecules made by piecing together DNA from different
organismsThis 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 vesselsmarketed 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 bacteriaCan be supplemented in individuals that are
deficient in a protein, like InsulinCan be made cheaper and safer than other
methods
Making a Recombinant DNA Molecule Step 1: Isolation of human DNA encoding
Insulin: Isolate Human DNACut 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 vectorCloning 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 plasmidCut 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
TThinkhink-P-Pairair-S-SharehareWhy is the plasmid with the
human insulin DNA considered a recombinant DNA molecule or chimera?
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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
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TThinkhink-P-Pairair-S-SharehareIYOW define transformation
of bacteria.
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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 antibioticAntibiotic –chemicals that prevent the growth
and survival of bacteria How does the antibiotic work?
Definition
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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
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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
TThinkhink-P-Pairair-S-SharehareYou 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.
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Uses of Biotechnology in Plants
Make Transgenic Plants toDelay 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 useHormonesBlood clotting factorsVaccines
Q & A: Recombinant DNA1) 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?