From Prokaryotes to Eukaryotes: Solving Problems Using Plasmids

Materials Needed

Activity 1 and 2:

• Interactive notebooks or 1 sheet of blank paper per student
• Prokaryote vs Eukaryote Interactive Notebook Cutouts, 1 copy per student

Activity 3:

• Plasmid DNA packet, 1 copy per group of 2-3 students
• Plasmid Problem Solving Scenario, project digitally for class to see
• Scissors
• Clear tape
• Three different colored pencils or highlighters

Essential Files (maps, charts, pictures, or documents)

• Plasmid DNA packet
• Plasmid Problem Solving Scenario
• Prokaryote vs Eukaryote Interactive Notebook Cutouts

Vocabulary Words

Background

Interest Approach - Engagement

1. Display the crop collage picture for students to see. Ask, “What do these 10 crops have in common?” If needed, identify each crop from left to right as corn, soybeans, cotton, canola, sugar beets, alfalfa, papayas, squash, apples, and potatoes. After students offer their ideas, tell them that each of these 10 crops has at least one variety that contains a gene from another genome. These are most of the crops in the United States that have a commercially available GMO variety available for farmers to grow.
2. Draw on students’ prior knowledge of genetics reminding them that each genetic trait possessed by an organism is coded in their DNA. Review examples of traits determined by genetics. Examples of genetic traits in plants could include flower or leaf color, size, resistance to disease, insect resistance, herbicide tolerance, etc.
3. Provide further examples by referring back to the GMO crop collage. Point to the corn, soybean, and cotton plants and explain that the GM variety of these crops has a gene making it resistant to specific insects that kill or damage the plant. Without spraying insecticide, the plants are resistant to insects that could otherwise destroy the crop. Next, point to the corn, canola, alfalfa, soybean, cotton, and sugarbeet. These six crops have a GM variety that is resistant to a specific type of herbicide (chemical used to kill weeds.) This allows farmers to spray the plants with an herbicide to kill the weeds (unwanted plants) without killing the crop. Last, point to the squash and the papaya. These two crops are resistant to specific plant diseases.
4. Summarize by drawing a simple DNA double helix on the board. Point out that every genetic trait (such as the ones we just pointed out) an organism possesses is found in a section of its DNA structure. Using biotechnology, these useful genes can be identified and transferred to benefit other organisms. In these examples, the genes benefit plants used for our food supply, to feed livestock that provide us with meat, eggs, and milk, and to produce the fiber (cotton) we use for our clothes.
5. Ask students, “How is a desired gene transferred from one organism to another?” Tell your students that the answer is in a specific kind of cell structure and they will be finding out soon. Image source www.gmoanswers.com

Procedures

 Activity 1: Comparing Cell Types

1. Ask students, “Where are cells found?” Students should recall from their prior knowledge that all living things are made up of cells. They are found in anything living, including plants of all types, animals, and humans.
2. Tell students that there are two major types of cells. They are called prokaryotes and eukaryotes.
3. Give each student 1 sheet of blank paper or have them open to a new page of their interactive notebooks. Also, give them 1 copy of the attached Prokaryote vs Eukaryote Interactive Notebook Cutouts.
4. Instruct students to cut out the layered circle tabs and the diagrams of each cell type. This step may also be completed as bell-work as students come into class to save time.
5. Once the notebook page/worksheet has been assembled, review it as a class and discuss the differences and similarities of each type of cell. Use the following questions as a formative assessment:
   • Which type of cell is relatively larger than the other and usually part of a multicellular organism? (Eukaryote)
   • Which type of cell has a membrane-bound nucleus and organelles? (Eukaryote)
   • Which type of cell is usually a bacteria and is responsible for strep throat? (Prokaryote)
   • Which type of cell has DNA in a circular structure called a plasmid? (Prokaryote)
   • Which type of cell has DNA that is linear and found mostly in the nucleus? (Eukaryote)
   • Which type of cell makes up the structure of a plant or animal? (Eukaryote)
   • Which type of cell divides through mitosis or meiosis? (Eukaryote) 

Activity 2: What is a plasmid?

1. Ask students to look closely at the diagrams of their eukaryotic and prokaryotic cell from Activity 1. Ask, “ Is there a structure in the prokaryotic cell that you have not seen or heard of before?” Students may identify the flagellum or pilus. These answers are correct, but have them keep looking. When they identify the plasmid, tell them that they have found the cell structure that was an important tool in creating the genetically modified plants you introduced them to in the beginning of the lesson.
2. Have students place and label the plasmid diagram (from the printout used in Activity 1) on their notes page and write, “What is unique about a plasmid?”
3. Drawing on student’s knowledge of chromosomal DNA, discuss the unique characteristics of a plasmid and the DNA it contains.

Activity 3: Plasmid Problem Solving

1. Divide students into groups of 2-3, and give each group one copy of the Plasmid DNA packet, scissors, clear tape, and three different colored pencils or highlighters.
2. Instruct students to cut out the strips from page 1 of their packet (titled Plasmid DNA) and tape them consecutively into one, continuous strip (first strip 1, then strip 2, etc.). Then, tape the end of strip 6 to the beginning of strip 1 to make a ring. Check to be sure that none of the bases were covered up in the process of taping. (Pages 2 and 3 of the packet will be set aside until step 7)
3. Once the paper plasmid has been created, review with students what they learned about plasmids in Activity 1 and Activity 2. Ask questions such as:
   • What kind of cell would contain a plasmid? (prokaryote)
   • What is a plasmid composed of? (DNA)
   • What kind of traits does plasmid DNA typically code for? (traits giving the cell a selective advantage over others such as resistance to a strain of bacteria, fungi, or antibiotics.)
4. Instruct students to keep the last question in mind (plasmid DNA can code for resistance traits) as you introduce them to a scenario.
5. Project the attached Plasmid Problem Solving Scenario sheet for students to see. Ask students to brainstorm some solutions to this farmer’s problem. Ask, “How can the rootworm be controlled?” If time allows, give students a few minutes to research methods of rootworm control on their own. If time is short, inform students that farmers can use insecticides to kill the pest or they can rotate their crops from year to year to break the life cycle of the rootworm.
6. Ask students to think deeper about what they have learned about plasmids. Could science (specifically biotechnology) provide a solution? (Yes)
7. Ask students to find page 2 of their Plasmid DNA packet titled, “DNA Strip for Rootworm Resistance.” Instruct students to cut out the DNA strips and tape strip 1 to strip 2 (make sure students do not circle the DNA segment like they did the plasmid.) As students are cutting and preparing their DNA strip explain that this section of DNA came from a bacterial cell of the species Bacillus thuringiensis which we will call Bt for short. The shaded portion codes for the production of a protein that is toxic to corn rootworm larvae.
8. Inform students that their goal is to insert the shaded portion of DNA into the plasmid using the following steps:
   1. First, the plasmid has to be cut open. In it’s current circular shape, new DNA cannot be inserted. Ask, “How can a plasmid be cut?” (with a restriction enzyme) Prompt students to the correct answer by having them take a look at the remaining page (page 3) of their Plasmid DNA packet titled, “Restriction Enzymes.”
   2. Students should evaluate the available restriction enzymes to determine which enzyme is the best for this situation. The best restriction enzyme will be able to cut the plasmid, as well as the desired DNA strand in order to create “sticky ends” in the nucleotide base that can fit together like a puzzle. Direct students to review the restriction enzyme patterns within the cut area. Students should use three different colored pencils or highlighters to highlight the base pattern that matches enzyme cut area if the enzyme was used.
      • For example: Students will highlight CTAG in the enzyme Bam HI, GGCC in the enzyme Xma I, and CTG in the enzyme AVA II.
   3. Have students search the plasmid for the patterns highlighted in the restriction enzymes. Students should highlight these with corresponding colors. For example, if you highlighted CTAG in Bam HI green, highlight this base sequence in the plasmid green as well.
   4. Repeat the process in the DNA strand for rootworm resistance. After students have completed this step, ask which enzyme will cut both the plasmid and the DNA strand for rootworm resistance. (Xma I is the correct answer)
   5. Have students cut the shaded portion of the DNA strand at the cut sites for the enzyme.
   6. Open the plasmid at the enzyme cut sites and place the DNA strand inside the plasmid.
9. Once students have created their new plasmid with the desired (Bt) gene, explain that there are several steps left to create the transgenic plant. First, a step called transformation is needed to cox the bacteria to take up the newly transformed plasmid. By growing lots of the bacteria, scientists then have plenty of plasmid DNA to work with and can move on to the next step, getting the plasmid DNA to be incorporated into the corn plant's chromosomal DNA. There are many methods of doing this step. Once the plasmid DNA is part of the corn plant's DNA, the resultant corn crop will produce the Bt protein which is toxic to the rootworm and other damaging insects. The resistance trait will be passed from generation to generation with traditional plant breeding (cross pollination) and the rootworms will be controlled without the use of insecticide (a substance used to kill insects).

A common misconception is that GMO crops have increased the use of pesticides. It is important to understand that the word pesticide refers generally to substances used to kill harmful organisms. For example, herbicides kill selected unwanted plants and insecticides kill unwanted insects, but both are generally referred to as pesticides. As students come to understand the function of Bt corn they should recognize that it virtually eliminates the need for insecticides. When speaking specifically, the use of genetically modified Bt corn decreases the need for insecticides in crop production.

Concept Elaboration and Evaluation

After conducting these activities, review and summarize the following key concepts:

• Prokaryotic cells are smaller, contain smaller amounts of DNA, and are found in bacteria and protozoan.
• Eukaryotic cells are larger, contain more complex DNA (than prokaryotic) and are found in typical plant and animal cells.
• Plasmids are found in prokaryotic cells and are useful tools in biotechnology due to their containing smaller amounts of DNA compared to the DNA found in chromosomes and eukaryotic cells.
• Plasmids are a tool used in creating genetically modified organisms (GMOs).

Enriching Activities

• Assign students to work in groups to research one medical, agricultural and/or environmental use of biotechnology similar to what was demonstrated in Activity 3 of the lesson.
  •  Medical examples:
    • Pharmacogenomics is the study of testing the safety and impact of certain drugs based on the genetic information of the patient.
    • Gene therapy is used to integrate a beneficial gene into a patient in order to help cure a disease.
    • Insulin is made for diabetic patients using recombinant DNA technology. Scientists build the human insulin gene using bacterial plasmids.
  • Agricultural examples:
    • Corn, cotton, and potatoes have been genetically engineered to produce their own Bt (Bacillus thuringiensis). Bt is a naturally occurring bacterium that is found in the soil. The Bt in the plant allows the plant to be resistant to certain devastating pests.⁵
    • Some crops have also been genetically engineered to resist herbicides. This allows a farmer to spray herbicides without killing the plant. Glyphosate-resistant (GR) crops are a common type of herbicide resistant crop.⁶
  • Environmental:
    • Bioremediation is the process of using naturally occurring microorganisms —such as bacteria, fungi and yeast — to clean up polluted waterways, such as a body of water after an oil spill.⁷

Sources

Activity 3 was written by Heather McPherson and Mariel Sellers as part of the Syngenta Summer Fellowship Educator's Guide. The remaining activities were developed and added with permission by the National Center for Agricultural Literacy.

Author

Adapted from Plasmid Problem Solving found within the National Agricultural Literacy Curriculum Matrix. Original authors: Andrea Gardner, Heather McPherson, and Mariel Sellers

Organization Affiliation

National Center for Agricultural Literacy and Syngenta