Maximum distance between ATG & SP6 promoter

Nick Theodorakis nicholas_theodorakis at urmc.rochester.edu
Thu Nov 7 23:26:16 EST 2002


On 7 Nov 2002 06:36:20 -0800, rupertyip at msn.com (Studio401) wrote:


[...]

>
>We're trying to in vitro translate a cDNA using Promega's TNT kit. 
>Unfortunately, our cDNA was subcloned into a expression vector with
>~180 bp in the 5'-UTR upstream of the SP6 promoter.  I was just
>wondering if 180 bases was too far away for the SP6 promoter in this
>instance for expression.  Are there examples in the literature where
>people have expressed proteins where the ATG is =>180 bases away from
>the SP6 promoter?

Well, FWIW, once upon a time I used a beta-tubulin message with a
5'-utr of about that size that was synthesized by SP6  and it seemed
to translate fine in retic. lysate. 

Nick

-- 
Nick Theodorakis
nicholas_theodorakis at urmc.rochester.edu



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