Which strand does T7 RNA pol transcribe from?
brett at BORCIM.WUSTL.EDU
Mon Mar 10 09:37:55 EST 1997
>A simple enough question, but one which is causing me some bother!
> I have a double stranded DNA molecule about 150 bases in length.
>The problem is that the two strands are only complimentary for the
>first 60 or so bases. At the 5' end of the sense strand I have a T7
>RNA Pol promoter sequence. My question is: upon introduction of T7
>RNA Pol which strand is the template for the RNA Pol - the strand
>containing the promoter sequence or the strand containing the
>complement of the promoter sequence?
> My understanding of the situation is that the RNA pol uses the
>antisense strand as the template (i.e. the strand with the sequence
>complimentary to the promoter sequence). Therefore the rna generated
>will be the compliment of the antisense strand. Therefore when the
>sequences are no longer complimentary (after about 60 bases) the rna
>generated will continue to be the compliement of the antisense strand
>and will include the sequence unique to the antisense strand.
>Am i correct?
>Any help is appreciated.
You are correct. In fact, check out: Milligan, JF and Uhlenbehck, OC Methods
in Enzymology, V180, pp180-62. here they demonstrate transcription off the
Program in Immunology
Washington University - St Louis
brett at borcim.wustl.edu
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