<p><span style=\"font-family: Times-Roman;\"><span style=\"font-family: Times-Ro
ID: 11299 • Letter: #
Question
<p><span><span><p>53. You have just discovered a new life form from the ocean and named it</p>
</span><em><span>Spongeous maximus</span></em><span>. You want </span></span><span>to quickly check whether this organism carries a similar version (homolog) of a particular carrier protein
<p>that your lab has isolated and studied.</p>
<p>A) What critical information or reagent would you need in order to use GFP tagging to analyze this</p>
<p>protein?</p>
<p>B) What critical information or reagent would you need if you instead used a different technique that</p>
<p>involves either a western blot or microscopy?</p>
</span></p>
Explanation / Answer
Spongeous maximus!
A) What critical information or reagent would you need in order to use GFP tagging to analyze this protein?
Fluorescence tags are used to give visual readout on a protein. GFP and its variants are the most commonly used fluorescence tags. More advanced applications of GFP include using it as a folding reporter (fluorescent if folded, colorless if not).
You need flourescence tags and a lightbulb. You also need to know if
B) What critical information or reagent would you need if you instead used a different technique that involves either a western blot or microscopy?
Highly specific interaction of the ZZ-domain of Protein A, which constitutes part of the original TAP-tag, to rabbit IgGs can be used to detect TAP-tagged proteins in fixated cells by Confocal Laser Scanning Microscopy just by the use of labeled secondary antibodies. In addition, such interactions can be exploited for the analysis of transfected cells in FACS and Western blot experiments.
Bottom line: we need to know if this organism has antibodies (for part a) and secondary antibodies (part b).
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