Hi are you using mouse or human cells ? there is a good episomal systems for human cells: Grignani F, Kinsella T, Mencarelli A, Valtieri M, Riganelli D, Grignani F, Lanfrancone L, Peschle C, Nolan GP, Pelicci PG.High-efficiency gene transfer and selection of human hematopoietic progenitor cells with a hybrid EBV/retroviral vector expressingthe green fluorescence protein. Cancer Res. 1998 Jan 1;58(1):14-9. Kinsella TM, Nolan GP.Episomal vectors rapidly and stably produce high-titer recombinant retrovirus.Hum Gene Ther. 1996 Aug 1;7(12):1405-13. or you could use a retroviral vector. what is the problem for your system with making stable integrated lines using drug selection ? Good luck Rachel ======================================================= Rachel M. Gerstein, Ph.D. Department of Molecular Genetics and Microbiology Graduate Program in Immunology/Virology University of Massachusetts Medical School 55 Lake Avenue North Worcester, MA 01655-0002 (508) 856-1044 (508) 856-5920 (FAX) > ---------- > From: jack > Sent: Monday, March 25, 2002 1:20 PM > To: Cytometry Mailing List > Subject: Long-term GFP expression without selection > > Dear Flowers: > > Can anyone suggest a GFP vector which can stay stably in dividing cells > for a long time (such as 1 month) without G418 or other selection? We want > to try an in vivo study and need the information about the dynamic change > of targeted cell number. The GFP vector we tried is with CMV promoter and > the percentage of GFP(+) cells (PC-3) goes down very fast when cultured in > dishes without selection (even we cultured them from a single cell). Any > suggestion is deeply appreciated. > > Best Wishes, > > Jack > > >
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