I am trying to clone a cDNA encoding a extracellular protein. However, I
noticed that all of my positive clones (correct orientation) grew very
slowly and thus gave poor yields. I am using Bluescript. I suspect that
this sequence is toxic, and talking with a few others working with this
gene confirmed it may be. Now the question, what is toxic about it? Is
it the sequence of nucleotides, or is it producing some of the protein
accidently and this is toxic, or is it most likely an alternate open
reading frame coming from another cryptic promoter? I have heard all
three and was wondering if anyone could shed light on this. If it is any
of the above, does this mean that any DNA I recover is most likely
screwed up, rearranged, and would using ABLE or TOPP cells from
Stratagene help?
Tim Angelotti
TU Muenchen
Institute for Pharmakologie
