Message+to+Progen+1-5-12


 * from: |||| Kurt Whittemore kurtwhittemore@gmail.com ||
 * to: |||| info@progen.de ||
 * 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