C-terminal fusion vector

Iva Toudjarska toudjarska at wi.mit.edu
Tue Jul 16 15:29:37 EST 2002


Hi Frank,

There is system sold by NEB- IMPACT (Intein Mediated Purification with an Affinity
Chitin-binding Tag). The protein of interest can be cloned as a C- or N- terminal
fusion to a 55kD Intein-CBD. When you bind it to chitin column you can induce (with
DTT) the intein (protein splicing element) to separate the tag from the protein of
interest which you simply elute from the column.
Works well for some people.

Good luck
Iva

"Frank O. Fackelmayer" wrote:

> Hi all,
> does anyone out there know of and/or use a prokaryotic expression vector
> that allows cloning of a fusion construct with the fusion partner at the
> _carboxy_ terminus and the protein of interest at the amino terminus ?
> The fusion partner should be greater than 12kD in size.
> It seems there are only vectors available that are the other way round. Strange...
>
> Frank




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