Name Date Period

HBio

Jelly Genes Paper Plasmid Activity

Background

Certain jellyfish have the ability to glow (phosphoresce) and the gene for this ability is coded in the jellyfish genome. Can scientists, and indeed science students, insert this gene into other organisms? Today you will perform a transformation using a paper model.

What is a transformation? Bacteria have an extra piece of DNA that is much smaller than the rest of their genome, called a plasmid. This DNA is double-stranded and in the form of a circle. Transformation occurs when a gene from another organism is added to a bacterium’s plasmid, making that now part of the bacterium’s DNA. Can you think of some beneficial genes that could be added to bacteria? Remember that bacteria grow very rapidly, so they have the ability to make lots of protein in a short amount of time.

In the box below, write your understanding of the word transformation and then write your ideas for some beneficial genes that could be added to bacteria.

  • What is transformation?
  • What genes do you think could be added to a bacterium’s plasmid that would be beneficial to humans?

What are restriction enzymes and what do they have to do with transformation? In order to insert a gene into a plasmid, the gene must be cut out and an opening must be cut into the plasmid to give the gene a place in which to insert. This cutting is performed by molecular scissors, called restriction enzymes. These enzymes only cut in specific places. See the examples below and then complete the practice example.

Restriction Enzyme / Cut Site / Restriction Enzyme / Cut Site
EcoR / GAATTC
CTTAAG / XbaI / TCTAGA
AGATCT
HindIII / AAGCTT
TTCGAA / SmaI / CCCGGG
GGGCCC

Read through the following double stranded DNA sequence, looking for any HindIII and/or SmaIsites. Draw lines where these restriction enzymes would cut the DNA.

A G T G A C T C C C G G G T G A A T C A G A A G C T T C C T T A

T C A C T G A G G G C C C A C T T A G T C T T C G A A G G A A T

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Procedure

  1. Obtain the following materials: tape, scissors, plasmid DNA sheet (white), and jellyfish DNA sheet (green).
  2. Look through your bacterial plasmid DNA (white paper) for the EcoRI restriction enzyme recognition site. (See front of paper.)
  3. Draw a line depicting how the enzyme will cut the plasmid.
  4. Using scissors cut your plasmid where the EcoRI restriction enzyme would cut it. You are acting like a restriction enzyme.
  5. Look through your jellyfish DNA (green paper) for the EcoRI restriction enzyme recognition site. (HINT: You should find 2 recognition sites.)
  6. Draw a line depicting how the enzyme will cut the jellyfish DNA.
  7. Using scissors cut your jellyfish DNA where the EcoRI restriction enzyme would cut it. You are acting like a restriction enzyme.
  8. Recombine (or add) the jellyfish gene into the plasmid by matching the ‘sticky ends’ of the plasmid with the ‘sticky ends’ of the jellyfish gene.
  9. Using tape, tape the piece together. You are now acting as a ligase enzyme, taping the pieces together.
  10. Write your name on the green section and turn it after you have answered the questions below.

Conclusion

  1. Explain in detail what you have at the end of this paper simulation.
  1. Outline the steps (see diagram ) of transformation.
  1. Why might ‘sticky ends’ be better than ‘blunt ends’ when performing this type of genetic engineering?
  1. What could this type of genetic engineering be used for in real life?