How to detect GFP via FACS?

David N. Levy David_Levy at DFCI.harvard.edu
Thu Aug 17 10:15:55 EST 1995


In article <404v6r$ebs at bisance.citi2.fr>
Laurentiu COCEA <cocea at citi2.fr> writes:

> Apparently, someone has worked out a solution by mutating
> the original DNA sequence in order to have a different codon usage
> and managed to express the protein in mammal cells. But who, who?

See a scientific correspondence in Nature Vol 373 pg. 663-4 for the
article you ar looking for.  BTW, I have successfully expressed the
original GFP in a variety of fibroblastic and lymphoid cell lines.  I
PCR amplified only the open reading frame (there may be inhibitory
sequences in the 5' UTR) and added a Kozak translational start seqence
in the 5' primer, then cloned into a CMV promoter vector.  All these
factors seem to be involved in getting expression in lymhoid cells.  I
have not tried the improved version of GFP described in the article,
but it looks quite good.

David N. Levy, david_levy at dfci.harvard.edu
Dana-Farber Cancer Institute, Boston, MA



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