looking for a difficult gene - (Dec/07/2012 )
I am trying to carachterize a gene that has been sequenced in related species, but there is no way to find it in my species. I have tried Genome Walker, RACE and even tried at the protein level with immunoprecipitation and western blot with other species antibody. And the sriking point is that I always find related genes to the one I am looking for, and transposons, always of the same kind, but not my gene. Do you have any idea of what is happening, does someone has a similar experience?
Thanks in advance!
If there are regions of the gene that are strongly conserved, then it is often possible to design degenerate primers to pcr amplify a portion of the gene. Once you have that, then inverse PCR can give you the entire gene.
Here in the future, simply sequencing the organism would be another possibility.