Expression using metallothonein promoter in CHO cells
Harry.Witchel at Bristol.Ac.Uk
Fri Nov 1 07:58:12 EST 1996
Hallo Hallo Kings of DNA --
I have had inconsistent expression of a K+ channel driven by
a metallothonein promoter which is in a vector which also has hygromycin
resistance driven by a cytomegalovirus promoter. The cells are screened
for a month using hygromycin, so all the cells should have the plasmid by
now. When I add 10 uM Cd2+ to cells (CHO) grown on glass coverslips
(using Ham's F12 medium plus 10% FCS) and let expression go for 48 hours,
sometimes I get expression of my K+ channels, but often I don't. Also, I
find that my cells get very sick in the Cd2+, and they often round up,
and while patching they easily explode.
Is there a better way to induce the metallothionein promoter?
Is there a better inducible promoter (ie another vector)?
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