QUESTION IMAGE
Question
- what is the purpose of including the ampⁿ gene on the plasmid inserted into e. coli? (1pt)
- explain one reason why bacteria do transformation in natural settings. (1pt)
- fill in the expected results of the transformation of e. coli lab in the table below (4pts):
| lb plates | will e. coli grow? y/n | explanation for the result on the plate | will e. coli glow? y/n | explanation for the result on the plate |
| -dna | ||||
| -dna/+amp | ||||
| +dna/+amp | ||||
| +dna/+amp/+iptg |
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Brief Explanations
- ### Question 4
The $Amp^R$ gene encodes beta-lactamase, an enzyme that breaks down ampicillin. It acts as a selectable marker to identify successfully transformed E. coli that have taken up the plasmid, as only these bacteria can grow on ampicillin-containing media.
- ### Question 5
Natural bacterial transformation allows bacteria to take up free DNA from dead neighboring bacteria to acquire beneficial genes, such as antibiotic resistance genes, to survive changing environmental conditions.
- ### Question 6
| LB Plates | Will E. coli GROW? Y/N | Explanation for the result on the plate | Will E. coli GLOW? Y/N | Explanation for the result on the plate |
|---|---|---|---|---|
| -DNA/+Amp | N | Non-transformed E. coli do not have the $Amp^R$ gene, so they cannot survive exposure to ampicillin. | N | No bacteria grow on this plate to produce fluorescence. |
| +DNA/+Amp | Y | Only successfully transformed E. coli with the plasmid (carrying $Amp^R$) can grow in the presence of ampicillin. | N | IPTG is not present to induce expression of the GFP gene under the T7 promoter. |
| +DNA/+Amp/+IPTG | Y | Transformed E. coli with $Amp^R$ survive ampicillin and grow. | Y | IPTG binds to the lac repressor to induce transcription of the GFP gene, so the bacteria produce fluorescent GFP protein. |
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- Question 4: It is a selectable marker to identify transformed E. coli that can survive ampicillin treatment.
- Question 5: Bacteria use natural transformation to acquire beneficial genes (like antibiotic resistance) to survive harsh environments.
- Question 6: Completed table as shown in the explanations section.