1. Recombinant DNA Technology
Biotechnology
Recombinant DNA Technology
6/1/2018

2. Bellwork
Explain what genetic engineering means to you?
6/1/2018

3. Objectives Define recombinant DNA
Create a paper model of a recombinant plasmid and identify the key components
6/1/2018

4. Key Terminology Recombinant DNA molecule Chimera DNA ligase Plasmid
Vector
Transformation
Chromosomal DNA

5. Overview
Recombinant DNA Technology involves using methods to create new DNA molecules
New DNA molecules made by piecing together DNA from different organisms
This recombinant DNA (rDNA) is known as a chimera
Goat boy

6. Examples of Recombinant DNA Technology
rhInsulin, marketed by Eli, Lilly and Company
Recombinant human growth hormone (Nutropin, marketed by Genentech, Inc)
Recombinant “activase” (marketed Genentech, Inc)-dissolves blockages in blood vessels

7. Significance of rDNA
These proteins normally are produced in very small amounts or are synthesized at the wrong time in the human body
By being able to make these products in large amounts in other organisms, like bacteria.
Can be used to help individuals that are deficient in these proteins

8. Plasmid DNA
A small circular double stranded DNA found in bacteria and yeast
Separate from the chromosomal DNA
Chromosomal DNA - controls growth and division of organism

9. Characteristics of Plasmid DNA
Not involved in regulating growth and division of organism.
Consider autonomous in that it has control over its replication
Presence of plasmids in bacteria often present to protect against humans medicines by carrying antibiotic resistance genes
Prevents antibiotics from killing the bacteria

10. Why is a Plasmid called a Vector ?
Because the plasmid can replicate and carry recombinant DNA into organism of interest

11. Making a Recombinant DNA Molecule
Step 1: Isolation of human DNA encoding Insulin.
Isolate Human DNA
Cut DNA into fragments with restriction enzymes
Identify the DNA fragment that encodes for the protein, insulin

12. Making a Recombinant DNA Molecule
Step 2: Place DNA molecule into a cloning vector (commonly plasmid DNA)
Cut plasmid with same restriction enzyme used to make DNA fragments that contain insulin
Ligate fragments into the plasmid with DNA ligase
DNA ligase is an enzyme that “glues” the DNA fragments together to create the recombinant DNA molecule

13. Picture of Cloning Vector

14. Transforming Bacteria with Recombinant DNA
Step 3: The recombinant DNA molecule is taken up by the Bacterium by transformation
When a bacterial cell takes up foreign DNA (e.g. plasmid vector containing human insulin), it is said to be transformed
Bacteria then allowed to reproduce to make lots of cells containing the recombinant DNA molecule

15. PROPERTY OF PIMA COUNTY JTED, 2010
Think-Pair-Share
Why is the plasmid with the human insulin DNA considered to be a recombinant DNA molecule or chimera?
3. Think-pair-share
teacher presents a question
teacher gives wait time for student to form answer
teacher instructs students to share their answer with a partner
teacher calls on non-volunteers to share with the class
PROPERTY OF PIMA COUNTY JTED, 2010 3 15

16. Identify the Bacteria with the Recombinant DNA molecule
Must eliminate bacteria cells 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?

18. Antibiotic Resistance

19. Bacterium Takes Up Cloning Vector
Bacterium takes up cloning vector will now become ampicillin resistance.
In other words, will not be killed by the presence of ampicillin in the growth media
The bacteria without the cloning vector will be killed because do not contain the ampicillin resistant gene.

21. PROPERTY OF PIMA COUNTY JTED, 2010
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. Predit which petri plate will have more bacteria growth, the one + ampicillin or the one – ampicillin. Explain your reasoning.
3. Think-pair-share
teacher presents a question
teacher gives wait time for student to form answer
teacher instructs students to share their answer with a partner
teacher calls on non-volunteers to share with the class
PROPERTY OF PIMA COUNTY JTED, 2010 3 21

22. PROPERTY OF PIMA COUNTY JTED, 2010
12 Word Summary 48 37 18 21 14 13 23
In 12 words or less, list key concepts you learned today
Word Summary
in 12 words or less, summarize the most important aspects from today's lesson
teacher uses a strategy to check all
PROPERTY OF PIMA COUNTY JTED, 2010 22 22

23. Q & A: Recombinant DNA
Why is it useful to create bacteria that contain a new gene?
What is the purpose of restriction enzymes in this process?
What is the purpose of DNA Ligase?
Describe the 3 steps for making recombinant DNA.
Explain the specific steps you would use to create bacteria with a human growth factor gene.