Hi, Im wondering if mutagenesis typically works better with yeast haploids vs diploids (as will achieve more full knockouts etc)? The application is to generate increased production of a recombinant protein, not trying to characterise anything or do basic research. Currently using a UV procedure. Thanks
Generally speaking it would be easier to do this in haploids unless the gene of interest is essential. However if you are looking for gain of function changes then it might not matter.
Hi Jonathan,
If you do mutagenesis in a diploid strain, mutations carried by one chromosome, will be hidden by the homologue chromosome, and the phenotype will be similar to the WT, unless dominant mutations or genes giving a gene-dose response. As a consequence, mutagenesis in an haploid strain is more likely to provide you an answer. Best regards.
JL
Jonathan Bester
In your case choice of haploid strain is desirable. Since you have to increase the production of particular protein, and using random mutagenesis, it will work well and you will be able to see the better expected outcome with the haploid strain.
Jonathan Bester
There is no harm in using the diploid strain, you may just have to screen more colonies to get the desired mutant. For second question, I would recommend EMS and Concentration should be at least the same or more MIC90.- Badges
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