Message to Carlos at Scripps 1-5-12

2015-01-13

azim58 - Message to Carlos at Scripps 1-5-12

from:
 Kurt Whittemorekurtwhittemore@gmail.com

to:
 carlos@scripps.edu

date:
 Thu, Jan 5, 2012 at 5:40 PM

subject:
 pComb3XXS

Hi Carlos Barbas,

I am considering using your pComb3XXS vector to construct my phage
antibody library. I just had a few concerns/questions. I see that this
vector has the f1 ori. However, when I look at other phagemids, they have
the f1 ori and ColE1 ori both. I'm not sure why this vector does not have
the ColE1 ori. I don't understand the need for these oris clearly, but I
thought that the ColE1 ori helped to make sure that the plasmid would be
replicated when bacteria replicate.

Also, I see that this plasmid is designed for inserting the heavy chain
and light chain variable genes separately, and these genes will be
separated by a PelB Leader linker. However, before I started to consider
using this plasmid, I have already constructed scFv genes in the form of
heavy chain-linker-light chain where the linker is (Gly4Ser)3. Are there
any potential problems with me just inserting my scFv the way they are
now in place of the current heavy chain-linker-light chain in the
pComb3XXS vector?

Thanks for any information you can offer me! Feel free to educate me
about anything I am misunderstanding.

Best regards,
Kurt Whittemore

Graduate Student
Arizona State University
BIODESIGN INSTITUTE
Center for Innovations in Medicine
1001 S McALLISTER AVE
TEMPE, AZ 85287

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