Hello everyone!
I have tried use the Ig k signal sequence and PDGFR anchor sequence combination to express my target protein in HEK 293. But the target protein located almost all in nuclear membrane.
Do there have trick to construct cell surface expression constructs? or i have to screen different signal -anchor combination ?
Thanks !
Dear Liu-qing Chen
which is the origin of your sequence? (mammalian, bacterial, viral?) It is a soluble protein?
Sincerelly i never tried to do something like this, but for different purpose (screening the ability of mab to bind their antigens in HEK293 surface) i expressed several membrane antigens in the HEK293 surface.
I tihnk that possible solution is try to built a chimerae between a membrane protein that is generally well transfected in EXpi293 ( eg CD44, CCR8, or other) where you replace the protein N-term soluble part with your sequence, adding at the N-term the signal peptide of the membrane proteins.
If you proteins is not so big you can buy a sintetich construct cloned in pcdna3.4 from Genescript of other companies with some hundred dollars.
I'm not sure if ti will really work, but i'm optimist.
good luck
Manuele
Dear Manuele Martinelli
thanks for your helpful answer, i will try it. I also believe this method will work.
Sincerely
- Badges
- Science topic
- Similar topics
- Linguistics
- Composition Studies
- Publication