directional cloning - (Mar/20/2006 )
i wud like to clone a 3.4 kb gene into 5kb vector..
vector already contans promoter and ribosomal binding site...the sequences of promoter and ribosomal site were given...
my gene is in 5'to 3'direction...i.e sense strand or coding strand
which strand has to be ligated next to rbs(5'to 3'.....given on vector map ) to get the protein corresponding to the gene..
either sense or antisense?
can anybody help me..át really íi be appreciated.
The gene has to be transcribed 5'-3' with the translational "start site" closest to the promoter.
5'-Promoter -----gene------> terminator - 3'
Try to think of the gene as directional by understanding which way the gene is coded or transcribed. It can sometimes be confusing to think of the gene as sense and antisense, without understanding directionality.