YFP fussion protein problem - (Dec/04/2006 )
Hi everybody. I´ve got some strange results with a fussion protein with aminoterminal EnhancedYellowFP tag. The original overexpressed non-tagged protein yielded a diffuse nucleoplasmic localization by conventional immunofluorescence. However, the YFP-construct appears to accumulate in rounded, dense inclusions localized to nucleolus. Has anybody had any similar problem? Thanks in advance, have a nice day.
Miguelon
-miguelon-
I forgot to say that the YFP construct has been sequenced and is ok.
-miguelon-
QUOTE (miguelon @ Dec 4 2006, 02:52 PM)
Hi everybody. I´ve got some strange results with a fussion protein with aminoterminal EnhancedYellowFP tag. The original overexpressed non-tagged protein yielded a diffuse nucleoplasmic localization by conventional immunofluorescence. However, the YFP-construct appears to accumulate in rounded, dense inclusions localized to nucleolus. Has anybody had any similar problem? Thanks in advance, have a nice day.
Miguelon
Miguelon
it is known that original XFP tend to build at least dimers; to keep GFP monomeric A207 can be mutated to K; may also works for YFP
-The Bearer-