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Prop to Illustrate Lab 6: Bacterial Transformation Using Restriction Endonuclease
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by Shari Cohen
Hazel Crest, Illinois
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- One piece of colored material; should be about 80 cm long and 4 cm wide. This will represent the plasmid.
- Another piece of different colored material; should be about 250 cm long and 4 cm wide. This will represent a human chromosome (or any other organism's chromosome).
- Six to eight small pieces of Velcro.
Procedure
Put Velcro on the each end of the shorter colored material. Attach the two ends. This will represent the circular bacterial plasmid.Cut the longer material in three pieces. One piece can be 150 cm long, and the other two pieces about 50 cm. The exact measurements do not matter. Put Velcro on the ends of each of these pieces. Label one of the smaller pieces Gene A. Label the other small piece Gene B. Attach the three pieces together in any order. This will represent the long human chromosome.
Explanation
1. Create plasmid with the gene of interest: the gene of interest in this case is Gene A, which I will say codes for making insulin.
2. Plasmid is cleaved by restriction endonuclease.
3. Gene A will be cleaved by the same restriction endonuclease.
4. The DNA fragments with sticky ends (from plasmid and human chromosome) anneal and seal the sticky ends with DNA ligase.
5. Now the plasmid contains the gene of interest Gene A, insulin gene.
6. Transform recipient cells with plasmid DNA. The bacteria will reproduce the Gene A along with the bacterial genes.
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