Title: Quiz: pGLO Transformation
Course: BTEC-1000-301-302-303-F21 > Section: Genetic Engineering

Q: Which of the following best describes a plasmid?
- A small, circular piece of DNA that behaves like a minichromosome
- A large chromosome that carries most of the genes
- A gene that behaves like a protein
- A sugar that activates gene expression
A: A small, circular piece of DNA that behaves like a minichromosome

Q: The plasmid carries at least two genes.  One of those genes it the thing that you are interested in.  For example, it could be the gene for Green Fluorescent Protein or it might be the gene for insulin.  The other gene on the plasmid is called a "selectable marker".  What is the purpose of the "selectable marker" gene?
- It allows you to distinguish transformed cells from untransformed cells
- It allows you to select the time and place to express the gene of interest
- It allows you to carry out transformation much more quickly
A: It allows you to distinguish transformed cells from untransformed cells

Q: Will untransformed cells grow in the presence of ampicillin?
- NO
- YES
A: NO

Q: What is the effect of growing the cells in the presence of arabinose?
- The regulatory regions of DNA will be activated and the GFP gene will be expressed
- Large amounts of water will be absorbed by the cell
- The plasmid will be absorbed into the cells
- The cell wall will begin to degrade so that the GFP protein can be absorbed
A: The regulatory regions of DNA will be activated and the GFP gene will be expressed

Q: What does the word "transformation" mean?
- The genetic makeup of a cell is changed by the introduction of foreign DNA
- Green Fluorescent Protein is activated within the cell
- Bacteria are converted from a normal type to a laboratory type
A: The genetic makeup of a cell is changed by the introduction of foreign DNA

Q: You will start with two tubes labeled “+” and “-”. How are these tubes treated differently?
- The “+” tube gets plasmid DNA, while “-” does not.
- The “+” tube gets bacteria, while “-” does not.
- The “+” tube gets broth, while “-” does not.
- The “+” tube gets the heat shock, while “-” does not.
A: The “+” tube gets plasmid DNA, while “-” does not.

Q: How long should the heat shock last?
- 50 seconds
- 1 minute
- 5 minutes
- 10 minutes
A: 50 seconds

Q: After the heat shock step, we add broth to the cells and allow them to recover.  Why do we allow the cells to recover for a short time before streaking them onto the LB agar plates?
- To allow them time to make the antibiotic resistance protein
- To allow time for the plasmid to cross the cell membrane
- To allow the cells time to make cytochrome oxidase
- To allow time for the plasmid to supercoil
A: To allow them time to make the antibiotic resistance protein

Q: What is pGLO?
- Plasmid
- Protein
- Amino acid
- RNA
A: Plasmid

Q: What is GFP?
- Protein
- Plasmid
- Amino acid
- RNA
A: Protein

Q: What is a plasmid?
- A small circle of DNA.
- A DNA sequence that encodes a protein.
- A string of amino acids.
- A single stranded nucleic acid that is used to make proteins.
A: A small circle of DNA.

Q: In a successful pGLO transformation, the -pGLO LB/amp plate should not have any colonies. Why don’t the bacteria grow on -pGLO LB/amp?
- They have been killed by the ampicillin.
- They require the sugar arabinose to grow.
- They lack amino acids.
- They did not take up the GFP protein.
A: They have been killed by the ampicillin.

Q: In a successful pGLO transformation, the +pGLO LB/amp colonies should not glow green when exposed to UV light. Why don’t these colonies glow?
- They need the sugar arabinose to turn on the GFP gene.
- They have been killed by the ampicillin.
- They don’t have the necessary DNA to create the glow.
- They did not take up the GFP protein.
A: They need the sugar arabinose to turn on the GFP gene.