determining incorrectly spliced transcripts - (Apr/27/2002 )
Does any one know how to determine whether inserts in genes are relics of incorrect splicing of message?? if so has anyone studied what types of AA are likly to turn up in such an insert???? Any ideas greatly appreicated, cheers Dave.
I'd guess that incorrect splicing events are very rare, so if you can isolate the transcript several times from several animals/samples, then you'll have a reasonable indication as to whether your insert is an uncommon event, such as incorrect splicing, or a genuine variant form of splicing. Do you have genomic sequence available? That might help discriminate between splicing (variant or incorrect) and some other cause (maybe a random mutation in the cloning).
Hope this helps,
The expression of some genes is controlled by stocking the mRNA partially spliced in the nucleus. So the inserts that you observe may be the result of this type of control. To verify this hypothesis work only with material extracted from the cytosol.
helene, do you know how this sort of regulation works????? It sounds interesting.
It seems that the nuclear pre-mRNA splicing could impinge on three major aspects of gene expression: the production of differents protein isoforms, the kinetics of expression by altering the duration of the processing and the level of final product, though the efficiency of processing. I've read that src could act on pre-mRNA processing of TNFB by slowing down splicing and by acting on the export of partially spliced transcripts. For more information see Neel et al, current biology 1995,5(4):413-422.