pGLO Transformation Lab-25

pGlo Transformation Lab

​What is transformation?

What is genetic engineering?

​

The essential sequences in the pGLO plasmid include the following:

• GFP — jellyfish gene that encodes green fluorescent protein (GFP)

• ori — origin of pGLO plasmid DNA replication (essential for making more copies of the plasmid)

• bla — gene that encodes β-lactamase, an enzyme that breaks down the antibiotic ampicillin; transformants expressing the bla gene can be selected by adding ampicillin in the growth medium

The essential sequences in the pGLO plasmid also include the following:

• pBAD promoter — binds AraC-arabinose and promotes RNA polymerase binding and transcription of the GFP gene

• araC — gene that encodes a regulatory protein that binds to the pBAD promoter; only when arabinose binds to the AraC protein is the production of GFP switched ON

Multiple Choice

What are the important components of the pGLO plasmid?

• beta-lactamase gene, ribosomal RNA, terminator sequence
• araC gene, kanamycin resistance gene, promoter
• GFP gene, araC gene, beta-lactamase gene, and origin of replication.

• GFP gene, ampicillin gene, origin of replication

Multiple Choice

Why is the origin of replication an important component of the pGLO plasmid?

• The origin of replication allows the pGLO plasmid to be replicated within host cells.
• It is responsible for the color change in transformed bacteria.
• It determines the size of the plasmid.
• It helps in the transcription of genes on the plasmid.

Multiple Choice

What happens to gene expression when arabinose is NOT present?

• Gene expression is repressed.
• Gene expression is enhanced.
• Gene expression remains unchanged.
• Gene expression is activated.

​The pGLO plasmid sequence is below- what would happen if the highlighted base was deleted?

Multiple Choice

Which type of DNA mutation would cause a greater effect on the organism?

• Point mutation
• Silent mutation
• Inversion mutation
• Frameshift mutation

​LB -pGLO

​LB/amp
-pGLO

​LB/amp
+pGLO

​LB/amp/ara
+pGLO

Multiple Choice

The pGLO transformant bacteria in the LB/amp/ara environment glow under UV light because

• The bacteria express the green fluorescent protein (GFP) in the presence of arabinose.
• The bacteria are genetically modified to resist UV light.
• The bacteria glow due to the presence of ampicillin.
• The bacteria produce a red pigment in the presence of glucose.