"punctate" staining of GFP in E. coli

Paul N. Danese pdanese at BIOSUN.HARVARD.EDU
Sat Jan 25 23:02:41 EST 1997


I've been expressing a cytoplasmic GFP fusion protein in E. coli.  I had
assumed that this protein would be found evenly distributed throughout the
E. coli cytoplasm.  However, I observe several "dots" of fluorescence (2-11
dots) per cell, and I'm concerned that I'm concentrating fluorescent
protein into inclusion bodies.  I've typically read that GFP (and it's
derivative fusion proteins) is(are) not fluorescent in inclusion bodies.

However, has anyone ever observed FLUORESCENT GFP in inclusion bodies?

Has anyone ever expected to see even distribution of GFP within a cell and
observed localized fluorescence instead?

Thanks in advance for your help.


Sincerely,


Paul N. Danese
Department of Molecular and Cellular Biology
16 Divinity Avenue
Harvard University
Cambridge, MA 02138
email:  pdanese at biosun.harvard.edu
phone:  617-495-4217
FAX:  617-496-1114





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