C-terminal His tag

mj dreamwestus at yahoo.com
Wed May 14 15:56:27 EST 2003


Hi, all

I want to C-terminally His tag a protein from Caulobacter (a gram-negative
bacteria), and I am thinking about using pET-28a vector from Novagen.  But
this vector carries both an N-terminal His tag and an C-terminal one.  If I
clone my gene in frame with the C-terminal His tag, do I need to clone the
ribosome binding site of that gene from the Caulobacter (but I want to
express it in E.coli)??  The order of the sequence in that vector is: rbs--N
terminal his tag--restriction site--C termianl his tag
And do I need to remove the N-terminal his tag??

If I need to clone the rbs from the Caulobacter, is there any software or
tool that I can use to search for the ribosome binding site of a gene??  I
don't know the exact sequence of rbs in Caulobacter, and I think it's
different from E.coli

Help will be much appreciated!! Thanks!!






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