To identify the green fluorescent protein on the SDS-PAGE gel, we were trying to

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In tackling the question about identifying the green fluorescent protein (GFP) on an SDS-PAGE gel, we should evaluate what each option would reveal about GFP.
Option 1: 'compare the bands from transformed and non-transformed bacteria' — This approach could help identify any new bands that appear in transformed samples, but it does not specifically confirm GFP identity. Other expressed proteins or differences between samples might confound interpretation, and GFP’s fluorescence isn't assessed by just comparing band positions.
Option 2: 'compare the bands with a purified GFP standard' — Comparing to a purified GFP standard could help confirm the molecular weight and possibly the presence of GFP by size, but GFP is best confirmed by its intrinsic fluorescence rather than solely by co-migration with a standard. Gel mobility alone cannot guarantee correct identification, since other proteins might share similar sizes.
Option 3: 'check the gel for ethidium bromide fluorescence under UV light' — Ethidium bromide is a DNA stain; it fluoresces under UV light to visualize nucleic acids, not proteins. Using it here would not provide information about GFP, which is a protein and not stained by ethidium bromide in the same way.
Option 4: 'compare the bands from transformed bacteria cultured with or without arabinose' — This would address expression levels and inducibility of GFP under an inducible promoter, but it still does not directly identify GFP on the gel. It tells you whether GFP is produced under induction, not whether the band you see is GFP.
Option 5: 'check the gel for green fluorescence under UV light' — GFP has intrinsic fluorescence and will emit green light when illuminated with UV or blue light. Detecting this fluorescence on the gel directly confirms the presence of GFP in the sample, independent of protein size comparison or staining dyes.
Putting the options together, the reasoning hinges on whether the method directly signals GFP identity. Options 1, 2, 3, and 4 either rely on indirect inference, alternative stains, or expression criteria, rather than a direct test for GFP. Option 5 describes a direct fluorescence-based identification that matches how GFP is routinely detected in gels.
When considering the purpose of identifying GFP on an SDS-PAGE gel, the most straightforward and specific method is to observe green fluorescence under appropriate illumination, which aligns with GFP’s characteristic property. The other options provide useful context in some experimental designs but do not uniquely confirm GFP presence on a gel. The sequence of reasoning here helps disentangle why some approaches might seem reasonable but fail to specifically identify GFP, while the fluorescence-based method directly addresses GFP’s defining feature.

W25 BIOL D006B 01Y Cell & Molecular Biology Lab Quiz 4

To identify the green fluorescent protein on the SDS-PAGE gel, we were trying to

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