1. What is Irene’s problem?
Objective: Understand How Humans Benefit from Bacterial Transformation New Words: Insulin, recombinant DNA, plasmid, gene splicing
Irene is 10 years old and in the last few weeks, she suddenly experienced extreme tiredness, weight loss, and increased thirst. Her parents were concerned, so they took her to the doctor. Dr. Ross took Irene’s blood to test for her blood sugar. The results of the test indicated that she was a diabetic. When the doctor shared the results, Irene broke into tears. The combination of her sudden poor health and the news of her diabetes was too much for her to handle. “I don’t understand. What is diabetes and how can I get better?” The doctor felt bad for Irene. She said, “Irene, I can explain how this happened, and how we can fix it.”
What is Irene’s problem?
Irene is diabetic

2. Objective: Understand How Humans Benefit from Bacterial Transformation New Words: Insulin, recombinant DNA, plasmid, gene splicing
People with diabetes may not have enough insulin or may not be able to use it properly.
Insulin is a hormone that controls the level of blood sugar (also called glucose) in your body.
The sugar then builds up in the blood and overflows into the urine, passing out of your body unused. This deprives you of an important source of energy.
What is Diabetes?
A disease in which a person has high blood sugar, because the body does not produce enough insulin

3. How Can Irene Get Better?
Objective: Understand How Humans Benefit from Bacterial Transformation New Words: Insulin, recombinant DNA, plasmid, gene splicing
All people with type 1 diabetes, and some people with type 2, need to take insulin to help control their blood sugar levels.
Type 1 diabetes means your body doesn’t make any insulin.
Type 2 diabetes means your body either doesn’t make enough insulin or doesn’t use it properly.
The goal of taking insulin is to keep your blood sugar level in a normal range as much as possible so you’ll stay healthy.
Over time, high blood sugar levels can cause serious health problems such as blindness and kidney failure.
How Can Irene Get Better?
Irene needs to take insulin

4. Objective: Understand How Humans Benefit from Bacterial Transformation New Words: Insulin, recombinant DNA, plasmid, gene splicing
The first successful insulin preparations came from cows (and later pigs). In the 1980's technology had advanced to the point where we could make human insulin. The technology which made this approach possible was the development of recombinant DNA techniques. In simple terms, the human gene which codes for the insulin protein was cloned (copied) and then put inside of bacteria.
A number of tricks were performed on this gene to make the bacteria want to use it to constantly make insulin. Big vats of bacteria now make tons of human insulin. From this, pharmaceutical companies can isolate pure human insulin
How Can We Make Insulin?
The human gene which codes for the insulin protein was put inside of bacteria

5. Objective: Understand the Main Steps in Bacterial Transformation New Words: Insulin, recombinant DNA, plasmid, gene splicing, restriction enzyme
Step #1
Remove DNA from human cell.
Use restriction enzymes to cut a segment of DNA that contains a gene of interest, for example, the gene regulating insulin production.

6. Objective: Understand the Main Steps in Bacterial Transformation New Words: Insulin, recombinant DNA, plasmid, gene splicing, restriction enzyme
Step #2
Remove a plasmid from a bacterium and treated with the same restriction enzyme.
What is a plasmid?
A small, circular, DNA molecule present in Bacteria

7. Objective: Understand the Main Steps in Bacterial Transformation New Words: Insulin, recombinant DNA, plasmid, gene splicing, restriction enzyme
Step #3
Bind the insulin gene with the opened plasmid to form a recombinant plasmid.

8. Objective: Understand the Main Steps in Bacterial Transformation New Words: Insulin, recombinant DNA, plasmid, gene splicing, restriction enzyme
Step #4
The recombinant plasmid is re-inserted back into the bacterium.

9. Objective: Understand the Main Steps in Bacterial Transformation New Words: Insulin, recombinant DNA, plasmid, gene splicing, restriction enzyme
Step #5:
Bacteria clone into a large number of identical daughter cells

10. Objective: Understand How Humans Benefit from Bacterial Transformation New Words: Insulin, recombinant DNA, plasmid, gene splicing
Step #6:
The recombinant plasmid replicates as part of the bacteria’s DNA.

11. Objective: Understand How Humans Benefit from Bacterial Transformation New Words: Insulin, recombinant DNA, plasmid, gene splicing
Step #7
Insulin is produced by bacteria and extracted for human use.

12. Figure 13-9 Making Recombinant DNA
Objective: Understand How Humans Benefit from Bacterial Transformation New Words: Insulin, recombinant DNA, plasmid, gene splicing
Figure 13-9 Making Recombinant DNA
Gene for human insulin Recombinant DNA
Gene for human insulin Sticky ends
Bacterial cell containing gene for human insulin Plasmid
DNA recombination DNA insertion
Gene for human insulin
Recombinant DNA
Gene for human insulin
DNA recombination
Human Cell
Sticky ends
DNA insertion
Bacterial Cell
Bacterial chromosome
Bacterial cell containing gene for human insulin
Plasmid

13. Objective: Understand How Humans Benefit from Bacterial Transformation New Words: Insulin, recombinant DNA, plasmid, gene splicing 8
Bacteria clone into a large number of identical daughter cells
Bind the insulin gene with the opened plasmid to form a recombinant plasmid.
Extract the DNA from a human cell.
Insulin is produced by bacteria and extracted for human use.
Remove the plasmid from a bacterium.
The recombinant plasmid is re-inserted back into the bacterium.
The recombinant plasmid replicates as part of the bacteria’s DNA.
Treat the bacterial plasmid with the same restriction enzyme.
Use restriction enzymes to isolate a segment of (DNA) that contains the insulin gene. 5 1 9 3 6 7 4 2

14. Objective: Understand the Main Steps in Bacterial Transformation New Words: Insulin, recombinant DNA, plasmid, gene splicing, restriction enzyme
Describe the picture above following the numbered sequence.
Assure to include these terms: restriction enzymes, DNA, plasmid, gene, recombinant plasmid, replication, cloning, insulin.

15. Objective: Understand the Main Steps in Bacterial Transformation New Words: Insulin, recombinant DNA, plasmid, gene splicing, restriction enzyme
Step #1
Extract the DNA from a human cell
Use restriction enzymes to isolate a segment of (DNA) that contains the insulin gene

16. Objective: Understand the Main Steps in Bacterial Transformation New Words: Insulin, recombinant DNA, plasmid, gene splicing, restriction enzyme
Step #2
Remove the plasmid from a bacterium
Treat the bacterial plasmid with the same restriction enzyme
What is a plasmid?
A small, circular, DNA molecule present in Bacteria

17. Objective: Understand the Main Steps in Bacterial Transformation New Words: Insulin, recombinant DNA, plasmid, gene splicing, restriction enzyme
Step #3
Bind the insulin gene with the opened plasmid to form a recombinant plasmid.

18. Objective: Understand the Main Steps in Bacterial Transformation New Words: Insulin, recombinant DNA, plasmid, gene splicing, restriction enzyme
Step #3
The recombinant plasmid is re-inserted back into the bacterium

19. Objective: Understand the Main Steps in Bacterial Transformation New Words: Insulin, recombinant DNA, plasmid, gene splicing, restriction enzyme
Step #4:
Bacteria clone into a large number of identical daughter cells .

20. Objective: Understand How Humans Benefit from Bacterial Transformation New Words: Insulin, recombinant DNA, plasmid, gene splicing
Step #4:
The recombinant plasmid replicates as part of the bacteria’s DNA.

21. Objective: Understand How Humans Benefit from Bacterial Transformation New Words: Insulin, recombinant DNA, plasmid, gene splicing
Step #4
Insulin is produced by bacteria and extracted for human use.

22. Objective: Understand How Scientists Manipulate DNA New Words: E
Objective: Understand How Scientists Manipulate DNA New Words: E. coli, Ampicillin, GFP, plasmid, antibiotic
Do Now: List 3 things you learned in the Harlem DNA Lab

23. Objective: Understand How Scientists Manipulate DNA New Words: E
Objective: Understand How Scientists Manipulate DNA New Words: E. coli, Ampicillin, GFP, plasmid, antibiotic.
Complete the table below:
+GFP
+GFP LB LB
LB/Amp
LB/Amp
Transformed bacteria
LB (food)
Ampicillin
GFP
What do you have in the dish?
What is your hypothesis?
What are the results?
How do you explain it?
Non transformed bacteria
LB (food)
Non transformed bacteria
LB (food)
Ampicillin
Transformed bacteria
LB (food)
GFP
Non transformed bacteria spread across the entire plate
All bacteria grow across the entire plate
Less green colonies
Transformed bacteria
grow in the form of green colonies
No growth
Hypothesis is true
Transformed bacteria
grow in the form of green colonies
Same as expected in hypothesis
Same as expected in hypothesis
Same as expected in hypothesis
Non transformed bacteria feed on LB
Nothing stop them from growing
Non transformed bacteria killed by antibiotic ampicillin
Non transformed and transform bacteria feed on LB
Nothing stop them from growing
Only transformed bacteria can survive and grow in the presence of antibiotic ampicillin