I would like to start working with the C. elegans genome. For my new
project I would have to clone some C. elegans genes. I have got some
general questions concerning that.
My aim is to amplify the desired genes via PCR from cosmid clones
containing genomic DNA of C. elegans, so starting with the ATG, ending
with the stop codon and including all the introns. Afterwards I have to
express the genes in mammalian cells. Now I am wondering about the
splicing in the mammalian system.
So first, is there something known about the compatibility between the
mammalian and the C. elegans system concerning splicing? Are the systems
this conserved that C. elegans pre-mRNA can be correctly spliced in the
mammalian cells? Do you know something about a study where the same
approach that I want to do has ever been done? Going through the
literature I only saw cloning strategies starting with cDNA.
Next I red something about trans splicing in C. elegans. Is it possible
that mRNAs created from the gDNA are not stable without carrying the
trans spliced leader sequence?
Thank you very much for your help.