microheterogeneity and tagged proteins

[Chris Larsen]

If you can get the gene into the pRSET (invitrogen) system, you can get about 5-35 % recombinant protein in the bug and dont NEED to use the tag.  All you need is a CATATG start site to clone into Nde I.  I have used this method to prpare enough protein to crystallize.

Additionally, the cleavage site for this tag is specific (Enterokinase at DDDDK'D ?) but getting complete cleavage is actually the problem.

Chris