promoters for macrophage lines

Guriq Basi gbasi at SLIP.NET
Wed Feb 19 20:44:25 EST 1997


Dear Dr. Wade,

just read your posting on immuno.net.bio.net.  we are obtaining good
expression levels in Thp1 cells with cmv promoter driven constructs. our
reporter gene is ß-gal.  we've also tested rsv-ltr, mmtv, and sv40 in
transients, none were as good as cmv.  we're transfecting by
electroporation using a biorad gene pulser, 10X10(superscript)6 cells,
20µg DNA, 220V/960µF.  

now a question for you.  i'm trying to obtain stably transfected thp1
cells. after 3 mos effort, i'm still looking for that g418 resistant
colony.  it seems my transfection is killing off the cells before the
selection.  have you made stably transfected thp1 lines, or heard of any
one who has succeeded.  any details or info will be greatly appreciated.
thank you.  sincerely,

guriq basi, Ph.D.
athena neurosciences.



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