Hi,
I wished to amplify the CFP sequence off of pPD134.96 (fire lab vector kit) to tag my protein of interest. In the plasmid details, it lists that the construct results in equal nuclear vs cytoplasmic distribution. I am a little confused since ideally I would want it to work such that it shows actual localization of my protein of interest and not some artifact of the tag.Has anyone worked with this or similar vector before and can provide some insights from their experience?
Thank you
dear joan,
a little farther down in the fire vector kit info there is this paragraph, which should answer your question:
Nucleus≈cytoplasm: The reporter in these construct consists of the GFP coding region with no specific localization
signal. If these reporters are used without any additional coding sequence, the result is a cytosol-filling
fluorescent signal. Most cells expressing the reporter will show fairly uniform nucleus+cytoplasmic fluorescence,
although some show slightly stronger fluorescence in nuclei.