Boosting protein expression in mammalian cells ?

Nick Theodorakis nicholas_theodorakis at urmc.rochester.edu
Tue Jun 29 15:48:03 EST 1999


In article <3.0.32.19990629112558.008c1950 at imm2.imm.uth.tmc.edu>,
  dhavilan at IMM2.IMM.UTH.TMC.EDU ("David L. Haviland, Ph.D.") wrote:
> At 15:59 6/29/99 GMT, Nick Theodorakis wrote:
> >In article <3778D9F0.8AAF332B at cajal.mbb.ki.se>,
> >  svend at cajal.mbb.ki.se wrote:
> >> Dear All,
> >>
> >> Rumours around this lab have it that there is a drug out there, which
> >> may enhance the yield of transient heterologue protein expression in
> >> mammalian cells, e.g. CHO cells ?
> >>
> >> Can anyone confirm this and even better, name the compound ?
> >
> >IIRC, butyrate enhances expression from some promoters (e.g., CMV
> >promoter) a few-fold in transfected cells. If nobody else remembers,
> >I'll try to come up with a reference for you.
>
> Nick:
>
> It is sodium butyrate.  You have to take butyric acid and titrate in NaOH
> to bring the pH to 7.2-7.4.    Gorman, NAR 11:7631 (1983) I think is what
> the other fellow is looking for.
>
> However, if memory serves, I thought sodium butaryate only works with SV40
> based promoters as described in the paper.  They only used vectors with
> SV40 promoters.
>
> I thought TPA (or PMA to some) was better for CMV based promoters,  but I
> could be mistaken.
>
> Hope this helps,
> David
> =============================
>  David L. Haviland, Ph.D.
>  Asst. Prof. Immunology
>  University of Texas - Houston, H.S.C.
>  Institute of Molecular Medicine
>  2121 W. Holcombe Blvd.
>  Houston, TX  77030
>  Internet:"dhavilan at imm2.imm.uth.tmc.edu"
>  Voice: 713.500.2413  FAX: 713.500.2424
>  ------------------------------------------------------
> The hardness of butter is directly proportional to the softness
> of the bread.
> =============================
>

I'm thinking of a paper by F. Cabral (senior author), two or three years
ago, in J. Cell. Biol (I think, or maybe MCB). They were overexpressing
alpha-tubulin in transfected CHO cells and boosting the expression with
sodium butyrate. I _think_ they were using the CMV promoter, but I would
have to look it up, and a lot of my stuff is still in boxes because I
just moved.

Nick

--
 _______________________________________________
| Nick Theodorakis                              |
| nicholas_theodorakis at urmc.rochester.edu       |
| (previously theodorn at gusun.georgetown.edu)    |


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