Chapter 14. Examining KDEL in a GFP Construct
Introduction
Analyze the Data
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Analyze the Data 14-2: Examining KDEL in a GFP Construct
You have genetically engineered a green fluorescent protein (GFP) construct containing a KDEL sequence. When the construct is transfected into normal human fibroblasts and examined using fluorescence microscopy, the fluorescence appears throughout the cytoplasm, as diagrammed below.
a. How would you explain this pattern given that KDEL is supposed to be an ER-specific sorting sequence?
It’s everywhere because GFP is not a secreted protein so it wouldn’t have a hydrophobic signal sequence to get it into the RER in the first place. The KDEL sequence would not be recognized because the protein itself is not in the right compartment.
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