pGLO Transformation Lab-25

What is genetic engineering?

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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

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

​LB -pGLO

​LB/amp
-pGLO

​LB/amp
+pGLO

​LB/amp/ara
+pGLO