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pGLO prelab 1. What does transformation mean? 2. What areas of biotechnology is transformation used? Give 2 examples. 3. Draw a picture of a transformed bacterial cell with the pGLO plasmid inside it. Include the substances that are in the surrounding agar plate. 4. What genes does the pGLO plasmid carry? What does each one do? 5. Why do scientists prefer to do transformation on a simple organism like bacteria? 6. Why do plasmids make good vectors (something that transfers DNA from one place to another)? Draw and label the 5 plates you poured for the pGLO lab. 7. On which of the plate would you expect to find bacteria most like the original non-transformed E. Coli colonies? Explain your answer. 8. If there are any genetically transformed bacterial cells, on which plate(s) would they most likely be located? Explain your predictions. 9. What other molecule is required for expression of the GFP gene? (hint look at plates 4 and 5) PGLO Results and Conclusions 1. Complete the following table by observing the 4 agar plates. Label Treatment Observations/draw -DNA/LB Bacteria has not had pGLO plasmid inserted. Its plated on normal luria broth agar with no ampicillin added. -DNA/LB/amp Bacteria has not had pGLO plasmid inserted. Its plated on normal luria broth agar with ampicillin. +DNA/LB/amp This bacteria has been transformed (carries pGLO plasmid). It is plated on luria broth with ampicillin. +DNA/LB/amp/ara This bacteria has been transformed (carries pGLO plasmid). It is plated on luria broth with ampicillin plus arabinose sugar. 2. Compare the –DNA/LB plate with the –DNA/LB/AMP plate. What can you conclude about normal E. coli? 3. Compare the –DNA/LB/Amp plate with the +DNA/LB/AMP plate. What can you conclude about the transformed E. coli? 4. Compare the –DNA/LB plate with the +DNA/LB/AMP plate. Why were the growth patterns different? 5. If +DNA/LB/AMP did not produce colonies, what could be the reason? 6. As you compare the original bacterial traits (-DNA) to the transformed bacterial traits (+DNA), what two new traits do the transformed bacterial cells now exhibit? 7. What kind of molecule is glowing?