I always have been curious how scientists build a vector. I intended to integrate certain pathway (with 5-7 enzymes involved) into S. boulardii., but need suggestion on what vector to use, promoters, terminators, selectable marker, etc.
I believe that you can just use any of the vast array of vectors from S. cerevisiae for your work in S. boulardii, so long as the selectable marker will function. There is already a literature on this yeast, see for example:
Metabolic Engineering of Probiotic Saccharomyces boulardii; http://aem.asm.org/content/82/8/2280.full
Hi, thank you Michael. I was actually referring to the fact that there is no available commercial auxotrophic mutant for Saccharomyces boulardii, to my best knowledge. If let say, I use a vector with URA gene as the selectable marker, then even if I were able to transform the yeast, I would not be able to select the successful transformant since the WT of S. boulardii has functional URA gene, too. In the paper you attached, the researchers perform some CRISPR-cas system to produce that mutant strain.
Yes, you would either need to obtain such a mutant strain or build one yourself. However there are also some yeast vectors with dominant antibiotic selectable markers such as G418 or hygromycin or bleomycin. There may be others as well.
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