There are two main ways to insert genes in yeast, either in a replicative plasmid or in an integrative plasmid. The latter takes advantage of the homologous recombination machinery of the host cell, however, the efficiency of integration can vary, among other things, due to the homologous arms flanking the target gene.
Thanks a lot! However, HR in S. cerevisiae is preferred over Non homologous recombination, unlike P. stipitis. That is why even with short flanking regions it is possible to achieve good levels of integration in the former yeast. I will go ahead and read the paper you attached. Thanks, again.
That has been a long time ago and maybe you already make your own research but you might use longer than Saccharomyces sequences. I generally used more than 500 bp homology. Read:
Deletion of the Pichia pastoris KU70 Homologue Facilitates Platform Strain Generation for Gene Expression and Synthetic Biology
Laura Näätsaari,
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3387205/
Good luck.
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