pGLO Plasmid Bacterial Transformation
Hard
Can you give bacteria a new trait by adding foreign DNA? A plasmid is a small ring of DNA. When it enters a bacterial cell, it changes what that cell can do.
You introduce the pGLO plasmid into E. coli. This plasmid carries two genes. One makes the bacteria resistant to the antibiotic ampicillin. The other produces a fluorescent protein activated by arabinose sugar.
Mix E. coli with cold calcium chloride and the plasmid DNA. Heat-shock the cells at 42 degrees Celsius for 50 seconds. Spread them onto agar plates with ampicillin and arabinose. After incubation, check your plates under a UV lamp. Transformed colonies will glow.
Hypothesis
The hypothesis is that the E. coli with the pGLO plasmid will be transformed into a ampicillin-resistant strain that produces fluorescent protein.
Method & Materials
You will label two microtubes, transfer cold CaCl2 into each tube, transfer a plasmid DNA solution to one tube, incubate the tubes, transfer the E. coli/pGLO suspension onto agar plates, and wrap the plates together and place them in an incubator.
You will need E. coli starter plate with large isolated colonies, LB broth 42C, water bath, CaCl2 tubes in ice, pGLO plasmid, foam rack for microtubes, UV lamp, sterile glass beads in a container, disinfectant solution for contaminated beads at side of room, pre-calibrated inoculation loops, and poured agar plates.
Results
The experiment results showed that the E. coli with the pGLO plasmid was transformed into a ampicillin-resistant strain that produces fluorescent protein. The most interesting observation was that the LB/amp/arab plate produced glowing colonies.
Why do this project?
This science project is interesting and unique because it demonstrates how a DNA plasmid can be used to transform bacteria into a ampicillin-resistant strain that produces fluorescent protein.
Also Consider
Experiment variations to consider include using different types of bacteria and different types of plasmids.
Full project details
Additional information and source material for this project are available below.Related videos
These videos explain the science behind this project and demonstrate key concepts used in the experiment.
Share this Science Project:
Related Science Fair Project Ideas
Expose colon cancer cells to three nicotine concentrations and use a colorimetric assay to see which ones multiply.
Hard
Build model peptides on a computer in three shapes and compare their energies to find out why nature prefers helices over flat chains.
Hard
Coat a vibrating quartz crystal with virus-catching antibodies and measure frequency changes to detect baculovirus particles.
Hard
Share this Science Project: