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Flashcards in pGLO- Lab32 Deck (13)
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1

What is the origin of replication do

Allows the plasmid to self replicate when the nuclear DNA replicates. Providing all progeny with a copy of the plasmid

2

What does the bla do

The marker gene that codes for beta-lactamase

3

What is beta-lactamase

An enzyme that breaks down beta-lactam antibiotics

4

What does the green fluorescent protein gene do

A marker Gene that codes for green fluorescent protein

5

What is a green fluorescent protein

The protein derived from jellyfish that fluoresces when exposed to UV light

6

What does the pBAD promoter do

The promoter sequence used by enzymes that break down the sugar abinose

7

Name all of the genes that are present on the engineered plasmid

Origin of replication (ori)
Bla gene
Green fluorescent protein gene (GFP)
pBAD promoter
araC gene
restriction sites

8

What does the araC gene do

Codes for DNA-binding protein called araC.

9

What is the araC protein

The off switch that blocks production of arabinose enzymes when arabinose is not present by preventing transcription of RNA from DNA at the promoter.

10

What does restriction sites do

Allow for insertion of foreign genes using restriction enzymes

11

How was E. coli introduced into the plasmid

Artificial transformation– Heat shock

12

Describe how to assess the bla gene

Grow bacteria on medium that contains ampicillin. If they grow colonies they have bla marker gene

13

Describe how to assess for the GFP gene

Grow bacteria on medium that contains arabinose . If bacteria fluoresces under UV light they have the GFP marker gene that's been turned on by presence of arabinose