 INTERACTIVITY

Transforming an Animal
How can you create a transgenic cow capable of producing human insulin in her milk?

Timing Chapter 16, Lesson 2

Part I: Making the vector

1. In order to create the transgenic cow, your lab first needs to create a DNA vector containing the
insulin gene. This step involves a considerable amount of scientific terminology. Make sure you
understand the meaning of key terms. Match the following terms with their correct definitions.

1. ampicillin resistance gene 5. restriction site

2. promoter 6. Origin of replication
3. Xhol 7. Ligase
4. exon
A(n) 4______ is a region of DNA that is not transcribed.
5
A(n) ______ is the location in the plasmid that is recognized by the restriction enzyme Xhol.
7
______ is an enzyme that joins DNA fragments together.
6
______ is the location on the plasmid where DNA replication begins.
2
A(n) ______ is a region of DNA that initiates transcription of a gene.
7
______ is a restriction enzyme that looks for the sequence TCGA.
1
The ______ is a gene that enables you to identify bacterial cells that have taken up the plasmid.

Part II: Making the DNA linear

2. Explain why you need to culture the E. coli bacteria on agar that has been infused with ampicillin.
By culturing E. coli on agar infused with ampicillin, only the bacteria containing the resistance gene can
grow, allowing for the isolation and growth of only genetically modified bacteria.

3. Discuss two reasons you need to cut the plasmids prior to inserting them into the fertilized egg.
Firstly, it allows for the removal of unnecessary or unwanted DNA sequences from the plasmid. Secondly,
cutting the plasmids at specific restriction enzyme sites creates sticky ends that can base-pair with the
complementary sticky ends of the cut DNA in the egg, improving the efficiency of the ligation process.

Unit 4  Genetics | Interactivity 103 Copyright © by Savvas Learning Company LLC. or its affiliates
Part III: Embryo Creation
4. Infer:  Explain why you may need to microinject insulin genes into many different eggs in order
to successfully create one transgenic cow.
Microinjecting insulin genes into eggs is a technique used to create transgenic cows that produce insulin in
their milk. However, not all eggs will successfully integrate the gene and produce a transgenic cow.
Therefore, many eggs need to be microinjected to increase the chances of producing a successful
transgenic cow.

Part IV: The Transgenic Cow

5. Summarize:  Three techniques can be used to confirm whether or not a given calf is transgenic.
Carefully read about these techniques in part IV of the accompanying SCO. Then describe the
techniques here using your own words.

PCR (Polymerase chain reaction)

PCR (Polymerase chain reaction) is a technique used to detect the presence of a specific DNA sequence,
such as the transgene, in a sample. A small piece of DNA is amplified using a specific set of primers that
bind to the transgene sequence. The amplified DNA can then be visualized to confirm the presence of the
transgene.

FISH (Fluorescence in situ Hybridization)

FISH (Fluorescence in situ Hybridization) is a technique that uses
fluorescent probes to detect the presence and location of a
specific DNA sequence within an intact chromosome. The
probes are labeled with a fluorescent dye that binds to the
transgene, allowing the transgene to be visualized within the

Protein Analysis (Western blotting)

Protein Analysis (Western blotting) is a technique used to detect the presence of a specific protein, such as
the insulin produced by the transgene, in a sample. Proteins are separated by size using gel
electrophoresis, and then transferred onto a membrane. The membrane is then probed with an antibody
that binds specifically to the protein of interest, allowing the presence of the protein to be detected.

Unit 4  Genetics | Interactivity 104 Copyright © by Savvas Learning Company LLC. or its affiliates