Message to Progen 1-5-12
2015-01-13azim58 - Message to Progen 1-5-12
from:
Kurt [email protected]
to:
[email protected]
date:
Thu, Jan 5, 2012 at 6:00 PM
subject:
pSEX81 Surface Expression Phagemid Vector
mailed-by:
gmail.com
Hi Progen representative,
I am considering using your pSEX81 vector to construct my phage antibody
library. However, before I considered using your vector I have already
constructed my scFv genes which consist of a heavy chain-linker-light
chain where the linker is (Gly4Ser)3. I see that the linker you typically
use in your plasmid is slightly different.
"The VH and VL genes were joined by a DNA-fragment coding for a flexible
18 amino acid residue linker containing the first six amino acids of the
CH1 constant region domain and the hydrophilic pig brain alpha-tubulin
peptide sequence EEGEFSEAR." Note that this linker is referred to as the
YoI tag.
Cloning procedure: To clone VH gene fragments the recognition sites of
the restriction endonucleases NcoI and HindIII are recommended. VL gene
fragments should be introduced by using the recognition sites of the
enzymes MluI and NotI.
I would like to clone my complete scFv with the linker that they already
have in place of the scFv stuffer that is already in the vector. Do you
think this should work okay or is it critical that I have the same linker
you recommend?
Thanks for any information you can offer me!
Best regards,
Kurt Whittemore
Graduate Student
Arizona State University
BIODESIGN INSTITUTE
Center for Innovations in Medicine
1001 S McALLISTER AVE
TEMPE, AZ 85287