cloning membrane protein with 2 tags - (Apr/04/2006 )
I want to clone the membrane protein with 2 tags (one in the middle of the protein and other on C terminus) for visualization.
How can I do this in simple way?
for c terminus, a PCR is ok.
but both tagging may be done in closely one step. First PCR of the first fragment. Fwd primer is classical and reverse primers contains a unique restiction site, the flag, and the annealing part.
Seond part of the gene is done by : fwd primer holds the unique restriction site inserted by the FLAG-rev primer and the reverse primer contains the second flag...
i've attached the picture
Thanks a lot. It looks pretty good.