I have recently cloned a PCR product obtained using yeast genomic DNA
as the template. The product was produced using degenerate primers,
designed to match sequences conserved across species in a particular
protein. Some of the sequence seems to match well with the expected
translated product, however, the clone does not appear to have a single
ORF (i.e., when translated in the 3 frames suggested by the assumed
orientation of the original primers, none of the 3 frames are a complete
One of the possibilities is that this is, indeed, the gene I'm looking for,
but it contains at least one intron.
I don't know much about introns in yeast nuclear genes. Basically I want
to know how to spot an intron from the primary sequence and where to find
basic info on yeast nuclear genomic introns and splicing.
I will post a summary of the best help I have received in response.
Genetics and Molecular Medicine