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orientation of promoter into viral construct


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#1 Bomber

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Posted 23 August 2011 - 01:48 AM

hey,

i would appreciate some opinion on the follwoing consideration -

i am sub-cloning an insert including an upstream promoter between the LTRs of a viral expression plasmid.
so - when i clone the gene of interest and its promoter, do i need to consider the orientation within this viral expression plasmid.
i am asking since the LTRs of the viral plasmid are also working as promoter - so I am wondering if there is interference in transcription??
any opinions appreciated!

best regards

#2 Discuss

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Posted 23 August 2011 - 10:47 PM

I think there would be interference in trancription. I doubt even if the orientation is changed, whether it will prevent interference since the 3'LTR can also act as a promoter (when 5' LTR is interrupted or something like that). I think any sequence between the LTRs would be replicated. Check the link below (Pages 386 and 387).

http://books.google....epage&q&f=false




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