EGFP fusion protein subcellular localisation?

Zhonglin Chai Zhonglin.Chai at med.monash.edu.au
Tue Dec 16 02:33:49 EST 1997


Dear netters,

        I am attempting to determine the subcellular localisation of a
novel protein in mammalian cells (COS, HeLa, etc). I have cloned my cDNA in
frame with EGFP using Clontech's pEGFP-c1 vector. The transfected COS cells
were green extensively, but more concentrating on nucleus. The problem was
the wild type EGFP (without cDNA insert) localised to nucleus too, that
made it difficult to interprete the result. Where does EGFP or GFP usually
reside in COS and HeLa cells? What vector system is the best to use for the
localisation purpose? All the comments and suggestions are appreciated.

Yours Sincerely



ZhongLin Chai, PhD
____________________________________________
Department of Pathology and Immunology
Monash University Medical School
Alfred Hospital
Commercial Rd, Prahran, VIC 3181, AUSTRALIA
Telephone: (61 3) 9276 2698 (lab)
           (61 3) 9276 2696 (office)
Fax:       (61 3) 9529 6484
email:     zhonglin.chai at med.monash.edu.au
____________________________________________





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