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Does the bacteria on the pGLO LB amp plate 3 contain the GFP gene?

Bio-Rad’s unique pGLO plasmid contains the gene for GFP and a gene for resistance to the antibiotic ampicillin. pGLO also incorporates a special gene regulation system that can be used to control expression of the fluorescent protein in transformed cells.

Does the pGLO plasmid fluoresce?

Bacteria transformed with the pGLO plasmid are selected by ampicillin resistance and when induced to express GFP, they glow fluorescent green under UV light!

Why is it possible for bacteria cells to express a gene that is originally found in jellyfish?

A specific species of jellyfish has a green fluorescent protein (GFP). Some cells can turn this protein on resulting in the glowing effect. This gene was inserted in the cells of bacterium, E. coli and when exposed to UV light, they emitted a similar green fluorescence just like on the jellyfish.

Can bacteria be genetically engineered to produce a jellyfish protein?

pGLO Bacterial Tranformation The pGLO plasmid has been designed to express green fluorescent pigment (GFP). The GFP gene was first isolated from jellyfish. With advancements in biotechnology, we can now transform E. Coli to express the GFP gene and glow under a green light.

Why does arabinose make the glowing show up?

Coli bacteria then retained the pGLO which was activated by an arabinase system. This basically means whenever arabinose is introduced to the bacteria, and in ideal situations, the bacteria should begin to glow.

Why is arabinose required to make the GFP glow?

The plasmid naturally contains an operon for arabinose digestion. if arabinose is present in nutrient medium, the operon’s structural genes for arabinose digestion will be turned on. Thus when arabinose is present to turn on the arabinose operon, GFP is produced and the bacteria can fluoresce.

What 3 genes does the pGLO plasmid contain?

This plasmid has been engineered to contain three core genes: the bla gene which encodes the enzyme β-lactamase, responsible for resistance toward the antibiotic ampicillin (AmpR); the green fluorescent protein (GFP) gene, originally derived from jellyfish (Aequorea victoria), which encodes the GFP; and, the arabinose …

What does M stand for in pGLOm?

DNA is TRANSCRIBED into mRNA, mRNA is TRANSLATED into a protein molecule. What is the part of the gene sequence that controls the transcription process? The Promoter. What does the “m” stand for in pGLOm? Mutation.

What gene is responsible for turning on the glow?

At the heart of the research is the jellyfish Aequorea victoria, whose genes instruct cells to produce a green fluorescent protein that is responsible for the glow. Insert the jellyfish gene successfully into another organism and its cells will produce the protein that stimulates the glow.

Why does the pGLO plasmid glow on an LB amp ARA plate?

The pGLO plasmid has the bla gene which allows for ampicillin resistance. There are glowing colonies surrounded by satellite colonies, because ARA turns on the araC gene which turns on the expression of the GFP gene. When this is expressed in the presence of UV light, it fluoresces.

What happens to the AraC protein when arabinose is present?

When arabinose is present, arabinose binds AraC and prevents it from interacting. This breaks the DNA loop. The two AraC-arabinose complexes bind to the araI site which promotes transcription. When arabinose is present, AraC acts as an activator and it builds a complex: AraC + arabinose.

What happens to AraC in the absence of arabinose?

In the absence of arabinose, the AraC protein assumes a different conformation and represses the ara operon by binding both to araI and to a second operator region, araO, thereby forming a loop (Figure 14-17b) that prevents transcription.

Under what condition is the AraC protein an activator?

arabinose

Under which conditions are the Lac structural genes expressed most efficiently?

Under which conditions are the lac structural genes expressed most efficiently? When glucose is absent and lactose levels are high, the lac structural genes are expressed the most efficiently. Without glucose, cAMP is produced and CAP can stimulate transcription of the structural genes.

Is arabinose a carbohydrate?

Arabinose is an aldopentose – a monosaccharide containing five carbon atoms, and including an aldehyde (CHO) functional group.

What is the difference between promoter and enhancer?

An enhancer is a sequence of DNA that functions to enhance transcription. A promoter is a sequence of DNA that initiates the process of transcription. A promoter has to be close to the gene that is being transcribed while an enhancer does not need to be close to the gene of interest.