basics about expression vectors - (Nov/29/2007 )
I'm totally new to expression vectors...
I will use one together with a reporter construct. I will overexpress an ligand activated TF and look for its activation on the reporter plasmid.
I guess I need to check that the protein is really being produced? I would like to use western but there is not any commersial antibody (fish) to use so then I have to order an ab produced just for me and this is a bit expensive. What is the common way to verify a working expression vector???? (And is it 'obligatory' or can I just hope that everything works with the reporter and thats it...)
And 1 more..
I have sequenced the whole coding sequence and som UTR, what should I put in the vector, just the sequence corr. from first to last aa or some more part of the 5' end??
I would really appreciate ANY advice :-)
Many people add a small epitope tag to the protein of interest in order to verify expression. Tagging is preferred when no antibody is commercially available.
Thank you Scolix!