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Hands-on Activity: Bacteria Transformation 

Contributed by: National Science Foundation GK-12 and Research Experience for Teachers (RET) Programs, University of Houston

Quick Look
Grade Level: 9 (9-12)
Time Required: 45 minutes
Expendable Cost/Group: US $0.20
Group Size: 2
Activity Dependency:
Introduction to Genetic Engineering and Its Applications
Subject Areas: Biology

Summary
Students construct paper recombinant plasmids to simulate the methods genetic engineers use to create modi ed bacteria. They learn what
role enzymes, DNA and genes play in the modi cation of organisms. For the particular model they work on, they isolate a mammal insulin gene
and combine it with a bacteria's gene sequence (plasmid DNA) for production of the protein insulin.
This engineering curriculum meets Next Generation Science Standards (NGSS).

Engineering Connection
Bacteria are the most common organisms modi ed by genetic engineers due to the simple structures of
bacteria cells compared to those of eukaryotic cells. Engineers are able to add genes to bacteria using
recombinant plasmids, which enable the bacteria to produce the desired bene cial proteins. Students use a
paper model to simulate this real-life process used by bio-technicians.

The Enterococcus faecalis bacterium lives

in the human gut.

Learning Objectives
After this activity, students should be able to:

Model and describe the process used by engineers to modify the genome of bacteria.
Explain why bacteria are genetically modi ed more often than other organisms.
Discuss possible applications for genetically modi ed bacteria.

Educational Standards
 NGSS: Next Generation Science Standards - Science
 International Technology and Engineering Educators Association - Technology
 State Standards

Materials List
Each group needs:
 scissors
tape, 3 small pieces
DNA Sequences for Cut-Outs, one per group; these represent plasmid DNA and mammal DNA containing the insulin gene; helpful to print
page 1 on di erent colored paper than page 2
Modeling Bacteria Transformation Worksheet, one per group
Assessment Questions, one per student
stapler (can be shared among groups)

Worksheets and Attachments

DNA Sequences for Cut-Outs (pdf)
DNA Sequences for Cut-Outs Answer Key (pdf)
Modeling Bacteria Transformation Worksheet (docx)
Modeling Bacteria Transformation Worksheet (pdf)
Modeling Bacteria Transformation Worksheet Answer Key (docx)
Modeling Bacteria Transformation Worksheet Answer Key (pdf)
Assessment Questions (docx)
Assessment Questions (pdf)
Assessment Questions Answer Key (docx)
Assessment Questions Answer Key (pdf)

Visit [www.teachengineering.org/activities/view/uoh_genetic_lesson01_activity1] to print or download.

Pre-Req Knowledge
A basic understanding of protein synthesis and DNA's role in the cell/body is helpful so students can follow how changes in DNA result in major
changes in the characteristics of organisms.

Introduction/Motivation
How big are bacteria? (Answer: most are only a few micrometers. Micrometers are one millionth of a meter. 1 cm = 10,000 micrometers; 1 mm =
1,000 micrometers) Bacteria are so small that most are less than one-tenth of the diameter of a human hair.

How many di erent species of bacteria exist? (Answer: Estimates vary from 10 million to 1 billion species.) So far, we have only discovered less
than 10,000 di erent species, but some scientists estimate that as many as 1 billion di erent species of bacteria may exist on Earth! How proli c
are bacteria? Bacteria are so plentiful that a 20-ounce bottle of water may contain up to 600 million bacteria!

Bacteria are everywhere, and most of the time they are harmless. In fact, many are bene cial to people because they are useful and necessary to
a healthy human body and environment. What are some examples of these "good" bacteria? (Possible answers: E.coli within the intestines of
mammals, bacteria within the soil, bacteria used to make foods such as yogurt.) Without bacteria, we would not be able to digest food or
produce some of our favorite foods such as yogurt and cheese. The bacteria we might generally call "bad" for us include those responsible for
causing illnesses like food poisoning.

Do you think genetic engineers could use and modify bacteria for any purpose? (Answer: Yes) Bacteria are the most commonly modi ed
organisms. Let's look at how we can modify these bacteria and why we would want to modify them.

Procedure
Background

Bacteria can be adapted to produce a number of useful materials. Because of the simple structures of bacterial cells, they are the most
commonly modi ed organisms. Many times, and in this activity, a gene is simply added to the bacteria, causing the bacteria to be able to
produce a useful protein, such as insulin. Other changes to bacteria can recon gure the cellular respiration product to create desirable
byproducts such as diesel or plastic molecules instead of the usual byproduct, such as carbon dioxide.

The common method used for genetically modifying bacteria is to use recombinant plasmids. Plasmids are circular pieces of DNA; when placed
near bacteria, the plasmid is absorbed and incorporated into the bacterial cell. Once inside the bacteria, the plasmid is treated the same as the
bacteria's original DNA. This means that the bacteria will use this new DNA from the plasmid to create proteins, and the plasmid will be
replicated when the cell divides.

The process of creating genetically modi ed bacteria used in this activity is one of the simplest methods. First, a desired gene must be selected
from some (any) organism, that is, the gene that codes for the creation of insulin protein, and removed from the DNA of that organism. Genes
are removed using restriction enzymes. These enzymes search for speci c nucleotide sequences in the DNA, called recognition sites, where they
"cut" the DNA by breaking certain bonds. When the bonds are broken in a staggered manner it creates "sticky ends." If the enzyme breaks the
bond to create a straight cut, the ends are called "blunt ends." The next step is to use the same restriction enzyme to cut open the plasmid.
The isolated gene is now placed where the plasmid was cut, and they are bonded together using another enzyme called ligase. Now a
recombinant plasmid has been produced. The nal step is to get the plasmid into a bacteria cell. Sometimes simply placing the bacteria in the
correct environment is enough to arti cially induce the bacteria to intake the recombinant DNA, otherwise some special method may be
required if the plasmid is too large to cross the bacteria's cell membrane. Figure 1.shows a diagram of the entire process.

Before the Activity

Gather materials and make copies of the Modeling Bacteria Transformation Worksheet, one per
group, and Assessment Questions, one per person.
Make copies of the DNA Sequences for Cut-Outs, one per group; it is helpful if the plasmid DNAs
(page 1) are printed on di erent colored paper from the mammal DNAs (page 2) to help distinguish
them during the activity. Then cut the DNA sequences into strips.
If time is short, tape the cut-out plasmid DNA into circles with the DNA sequence facing outward.
Otherwise, have students do this in step 2.

With the Students

1. Divide the class into groups of two students each. Hand out to each group a worksheet and its two
strips of DNA sequences, pointing out which will be used to create the plasmid, and which is the Figure 1. The process to build a recombinant
mammal DNA containing the insulin gene. plasmid to modify bacteria.

2. Direct groups to tape together the ends of the plasmid DNA to form a circular piece of DNA (see Figure 2) so that the printed sequence is
visible on the outside of the plasmid. This is the initial plasmid that will be modi ed with the insulin gene.
3. Find the recognition sites on both the plasmid and the mammal DNA. The restriction enzyme being
used will search for a speci c base pair sequence on the DNA to cut. This sequence and the cut
pattern are shown in Figure 3. Have students look for this sequence on both DNA strands.
Anywhere they nd the full sequence is a recognition site; have them draw a dotted line at the site
where the restriction enzyme will cut. Doing this usually prevents students from cutting straight
across in the next step.
4. Have students apply the restriction enzyme (represented by Figure 2. Illustration of the circular DNA created
scissors in this model) to cut the DNA at the marked in step 2.
locations. Performed correctly, students make one cut
through the plasmid and two cuts on the DNA containing the
insulin gene. These cuts are made on either side of the
designated gene. They are "staggered" cuts, forming the
Figure 3. A restriction enzyme recognition site
"sticky ends." Now that the desired gene is isolated, it is ready with dashed line showing where the enzyme will
to be added to the plasmid DNA (see Figure 4). break apart the DNA.

5. Next, have students use the ligase enzyme (represented by tape in this model) to rebuild the
plasmid with the new gene incorporated. If the DNA sequences have been correctly cut, the base
pairs from each end of the gene match exactly with the cuts made in the plasmid. Tape the ends of
the gene to the matching sticky end on the plasmid (see Figure 5).
6. If groups have correctly performed their genetic engineering
simulations, expect them to now have circular recombinant
plasmids that each contain the gene to produce insulin. Have
them staple their recombinant models on their worksheets
Figure 4. In blue, the model plasmid after being
and complete the worksheets as a team.
cut by a restriction enzyme. In red, the mammal
7. Conclude the activity by administering the Assessment DNA with cuts and its isolated insulin gene with
Figure 5. The nal recombinant model plasmid, a
Questions, as described in the Assessment section. Have sticky ends.
merger of the original plasmid DNA and the
students staple. added insulin gene.

Vocabulary/De nitions
DNA: Acronym for deoxyribonucleic acid, which is a molecule that contains an organism's complete genetic information.
Gene: The molecular unit of an organism that contains information for a speci c trait (speci c DNA sequence).
Genome: An entire set of genes for an organism.
Model: (noun) A representation of something for imitation, comparison or analysis, sometimes on a di erent scale. (verb) To make something
to help learn about something else that cannot be directly observed or experimented upon.
Plasmid: The circular DNA structure used by bacteria.
Prokaryote: A cell without a nucleus and membrane-bound organelles.
Recombinant DNA: DNA to which a section has been removed and replaced (recombined) with a new sequence.
Restriction Enzyme: An enzyme that "cuts" DNA when speci c base pair sequences are present.
Assessment
Pre-Activity Assessment

Review: Quickly do a review of bacteria, emphasizing bacteria structure. Discuss the pervasiveness of bacteria in everyday life and some roles
they play, both bene cial (digestive health) and harmful (illness). Alternatively, print out the Lesson Background section and have students read
through it.

Activity Embedded Assessment

Building a Model Recombinant Plasmid: Direct student pairs to apply their understanding of genetics to modify plasmids. After nishing the
modi cation, have teams complete the Modeling Bacteria Transformation Worksheet to demonstrate their understanding of the modi cation
process.

Post-Activity Assessment

Questions: Administer the Assessment Questions. Use the Assessment Questions Answer Key to individually gauge students' understanding of
the genetic modi cation of bacteria and its potential bene ts and dangers.

References
Bacteria. Updated December 9, 2013. Wikipedia, The Free Encyclopedia. Accessed December 10, 2013. http://en.wikipedia.org/wiki/Bacteria

Contributors
Matthew Zelisko, Kimberly Anderson

Copyright
© 2013 by Regents of the University of Colorado; original © 2013 University of Houston

Supporting Program
National Science Foundation GK-12 and Research Experience for Teachers (RET) Programs, University of Houston

Acknowledgements
Acknowledgements This digital library content was developed by the University of Houston's College of Engineering under National Science
Foundation GK-12 grant number DGE 0840889. However, these contents do not necessarily represent the policies of the NSF and you should not
assume endorsement by the federal government.

Last modi ed: June 20, 2019

Free K-12 standards-aligned STEM curriculum for educators everywhere.

Find more at TeachEngineering.org