How to link plasmid with peptide ?
klenchin at REMOVE_TO_REPLY.facstaff.wisc.edu
Mon Jan 29 19:24:57 EST 2001
In article <3A2038E37674D4119EA70050DA69A0C011BF1F at MMPS-PC-S6>, sidney at neuro.mpg.de (Sidney Cambridge) wrote:
>does anybody have any suggestions how I could link a peptide to a plasmid ?
>I've only heard so far that people coupled a peptide to polylysine which in
>turn bound the plasmid. I would prefer a more defined way but I can't come
>up with anything else.
>I would appreciate hearing your ideas - even the crazy ones :-) !
Well, DNA has NH2 and you can add cystein to the peptide's terminus.
Now all you have to do is to buy heterobifunctional crosslinker
(amine/thiol reactive), couple it first to DNA (since the peptide
might have amines too), quickly buffer exchange on spin gel
filtration column to remove non-reacted crosslinker, add peptide
and wait. Sounds quite simple to me.
Not sure how it's usually done. For many applications I imagine
that with the right DNA/peptide concentration found empirically
any crosslinker, even glutaraldehyde, will do.
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