Could someone help me to know if I am doing the right thing?
I am cloning a virus gene (cDNA) into a plasmid that would hopefully give vRNA upon transcription. I designed the construct so that the gene is flanked with polI polymerase promotor at 5' end (ATGgcctaa...) and polI terminator at 3' end. This would be sense cloning. Right?! Will this yield to transcription of viral RNA?
I got confused as I read all the posts on this topic. I know the RNA polymerase I transcribes the antisense DNA strand into RNA. But doest that mean I have to clone my gene different from what I described about to get it transcribed?That is should I design my restriction sites so that the inserted gene is flipped? e.g. polI promotor 3' (TACcggatt....) 5' pol I terminator?
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