Hi, I am aware that yeasts are eukaryotes and have the ability to do splicing such as in other eukaryotes. But I wonder, if let say, I want to express heterologous gene A in yeast, if I enter the whole native gene A sequence, how the yeast know where to cut--which are the introns and exons? On the other hand, if I insert only cDNA of gene A into yeast, would it be subjected to splicing--since they yeasts naturally do splicing, they may not know whether certain DNA has already been spliced or not? I hope someone could enlighten me on this as I am a newcomer in the field. Thank you in advance.
As far as I recall expression of heterologous genes from genomic sequence in yeast leads to abherent splicing of introns/exons and is not recommended (someone with up to date knowledge of yeast expression systems may be able to correct me on this). If you express a gene generated from cDNA none of the intron/exon boundary markers will be present anyway and these are highly conseved throughout eukaryotes so there shouldn't be a problem with further recombination.
if your purpose is to express heterologous gene, your construct should have a specific yeast recognizable promoter to drive the gene transcription. If that is the case, using the cDNA will be much more efficient to produce mRNA for protein translation rather than using native genomic DNA with that the splicing sites might not be recognized in your yeast strain.
Pre-mRNA of a heterologous gene can be process (spliced) in yeast. The paper mentioned below demonstrates a PLANT pre-mRNA were spliced in yeast cells.
Biochem Biophys Res Commun. 2002 May 17;293(4):1209-16.
Title: Plant pre-mRNA splicing in fission yeast, Schizosaccharomyces pombe.
https://www.ncbi.nlm.nih.gov/pubmed/12054504
**If you can access to this paper, please send me a copy. Appreciate it!!
Thank you all for the great share. They are very informative! I would need to read more on this. I have encountered papers where someone cloned some enzymes found in plant to yeast Saccharomyces boulardii and they only mentioned that they cloned gene xyz.. without specifically mentioning cDNA, so I assumed what they cloned is the whole gene.
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