Lesson #13 Problem set Genetic Engineering
Questions due Monday, April 16, 2012, 11:59pm
Genetics Agro 315
1. Below are steps performed to make an E. coli bacteria that can make
human insulin protein for the health care market.
a. Use heat shock or electroporation to get the recombinant human insulin
gene to go into a living E. coli cell.
b. Grow the E. coli in fermentation culture, extract and sell Humulin.
c. Modify the human gene and give it a promoter that will be turned on in E.
coli cells.
d. Extract the DNA from a human who does not have diabetes
e. Isolate and clone (make copies of) the gene encoding the insulin protein
List these steps in their proper order:_____________________________
View the “Who wants to be a genetic engineer” and “Gene Gun”
animations before doing #2 and #3.
2. Below are steps used performed to make Corn that makes a protein toxic
to European Corn Borers.
a. Culture Bt bacteria from the soil environment where they are found in
insect rich environments.
b. Cross the transgenic corn plants with other corn plants to pass on the Bt
transgene.
c. Coat copies of the modified Bt gene onto gold particles.
d. Grow corn plants from the “shot” corn cells
e. Extract the DNA from cultured Bt bacteria
f. Isolate and clone the insect toxin encoding gene from the Bt bacteria.
List these steps in their proper order:_____________________________
_____3. Assume you are a genetic engineer and want to make a transgenic rice
plant to have an enzyme that will make a vitamin produced in the kernel. You will
use a gene that encodes a protein from a daffodil and you want the gene
expressed at a high level in the rice seed. Place the following steps in the
correct order to genetically engineer the sorghum.
1. Insert the gene into a single cell of a rice plant.
2. Extract DNA from a daffodil plant
3. Clone the gene for daffodil vitamin synthesis
4. Design the gene to express itself at a high level in the seed while the
rice grain is developing
5. Breed a transgenic vitamin producing rice plant with a diseaseresistant rice plant.
List these steps in their proper order:_____________________________
_____4. For genetic engineering in plants to be successful, where should
scientists be focusing their transformation efforts?
a.) a single plant cell
b.) every cell of one seed
c.) every cell in one plant
d.) the leaves of the plant
View the “Agrobacterium” link and work through this animation.
5. List these steps in their proper order
____. Put leaf disks from tomato in the dish with the agrobacterium
____. Combine the Bt gene with the Ti plasmid (circle of DNA) vector
____. Transfer the tomato leaf disks into a Petri dish with plant cloning media
____. Clone a transgenic tomato plant that has herbicide resistance as a
selectable trait plus insect resistance from the Bt gene.
____. Apply herbicide to the plant cells as they are growing in the plant cloning
media
____. Introduce the Ti plasmid with the Bt gene into agrobacterium.
____ Place the agrobacterium into media plates where they can infect plant cells.
6. View the animation Gene Coding. Then your group should do the interactive
animation called “Transgene Design”. Answer the following questions on the
transgene design animation.
a. transgene #1 Event 176 Promoter from gene______________________
Coding region from gene__________________
b. transgene #2 Mon810 Promoter from gene______________________
and Bt11
Coding region from gene__________________
c. transgene #3 Mon863 Promoter from gene______________________
Coding region from gene__________________
d. transgene #1 Herculex Promoter from gene______________________
Coding region from gene__________________
7. . What is a selectable marker and how is this used in genetic engineering?
8. Describe two ways that Animal Genetic engineering is different than Plant
Genetic engineering.
9. Describe two steps that would be the same in Animal Genetic Engineering
and Plant Genetic Engineering.
10. Rank these five animals from easiest to genetically engineer (1) to most
difficult (4). Ties are not allowed.
___ elephant
___ salmon
___ mouse
___ cat
___ fruit fly
Describe your reasoning.
11. What type of animal transformation is most likely to give chimeric animals
(animals with some genetically engineered cells and some that are not)
___ pronuclear injection
___ blastocyst embryonic cell transfection
12. What type of animal transformation does not allow the animal genetic
engineer to practice some sort of cell level selection?
___ pronuclear injection
___ blastocyst embryonic cell transfection
13. Discussion Board question:
a. Animal or Microbe Genetic engineering project design. Draw a diagram below
that outlines the five major steps in a genetic engineering project. Your project
should be on an animal or bacteria that you would like to genetically transform so
that it contains a transgene which improves a trait in the organism or introduces a
new trait to the organism.
b. Share your genetic engineering plan with your group and then select one
project for your group. Improve and perfect the idea. Make a “poster” of your
group’s idea and we will display this poster and use it in future problem sets.
Your poster should include...
a. The motive for doing this is explained
b. All steps clearly shown including gene modification details and the
transformation method.
c. A timeline for the project.
14. Iron fortified transgenic plants!!
Assume you are a genetic engineer and want to make a transgenic plant that will
have more iron (Fe) in the part of the plant that people eat.
a. Choose the plant that will be your Fe fortified food:
__ carrot
___ potato
___ wheat
___ soybean
___ cantaloupe
b. Assume that you have patent rights to the following genes. Chose the gene
that will be useful for your project
___ root symplast gene: turned on in root cells
___ turnip starch synthetase gene: turned on in a turnip tap root
___ barley seed development gene: turned on in young seed cells of barley
___pumpkin fruit formation gene: turned on in a growing pumpkin
___ watermelon stem transporter: turned on in cells that ‘feed the phloem’.
___ pea cotyledon storage gene: turned on in pea cotyledons.
c. Explain why you chose the gene above.
d. Draw a diagram that describe your Fe Fortified Food GE project.
15. Which picture is the best depiction of a chromosome in the process of DNA
replication?
16. Complete the drawing of DNA replication by labeling the following in addition
to the three enzyme listed below.
Replication fork, leading strand, lagging strand, DNA polymerase I, ligase