orientation of promoter into viral construct - (Aug/23/2011 )
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!
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).