I constuct plasmid that contains FMD promoter, MOX terminator, HARS1, and URA3 gene from S. cerevisiae, then transform by LiAc method to H. polymorpha NCYC495 for three times. But every time don't have any colony.
Then, I try to find paper about using URA3 gene from S. cerevisiae as a selection marker in H. polymorpha. For H. polymorpha strain CBS4732 and DL-1 can use, but for strain NCYC495 don't have any paper report about ScURA3 gene.
Does anyone know that S. cerevisiae URA3 gene can function in H. polymorpha NCYC495 or not?
Any suggestion please provide
Hi there,
Yes! For info, read the following where ScURA3 cassette is used for gene disruption in your favorite host:
http://onlinelibrary.wiley.com/doi/10.1002/%28SICI%291097-0061%2819990930%2915:13%3C1323::AID-YEA459%3E3.0.CO;2-1/epdf
Dear Prof. Liger,
Now, I can't access to you link, but I will try with computer at university tomorrow.
Thank you
Hi,
You might want to check ur transformation process because the commonly used LiAc method for S. cerevisiae doesn't work in Pichia pastoris (a more closely related yeast to hansenula). Chemical method needs spheroplasting and use of LiCl. Mostly electroporation is used with Pichia
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