creating cell lines (both transient and constitutive)

Tom Anderson ucgatan at socrates-a.ucl.ac.uk
Mon Feb 23 06:39:26 EST 2004


On 21 Feb 2004, redeamer wrote:

> i need to do some cell lines (Huh7) expressing one protein or the
> whole RNA viral genome... but i have absolutely NO idea on how to do
> that sort of things.. Could anyone enlighten me on that subject...

You mean a bit like:

http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12482660&dopt=Abstract

I don't know if i've misunderstood your question, but in general, you make
a plasmid with your sequence of interest and a eukaryotic selectable
marker (like hygromycin resistance), transfect it into your cells somehow
(eg Novagen GeneJuice, Invitrogen Lipofectamine, old-fashioned calcium
phosphate, etc), wait a bit, then put on the appropriate selection and
wait for colonies to emerge. If you want transient expression rather than
actual cell lines, you leave out the selection stuff and just use a
transfection method.

HTH,
tom

-- 
Tom Anderson, MRC Laboratory for Molecular Cell Biology, UCL




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