Plasmid design
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We’ve chosen EcoRI and NotI as our restriction enzymes, because they cut only in the right places on our plasmid.
The following PDFs map the pPIC3.5K plasmid with each of the bovine casein genes inserted.
File:pPIC35K-Ost1-Alpha-AS1-sequence.pdf
File:pPIC35K-Ost1-Alpha-AS2-sequence.pdf
File:pPIC35K-Ost1-Beta-sequence.pdf
File:pPIC35K-Ost1-Kappa-sequence.pdf
We then used Benchling to optimize the codons for expression in the yeast Pichia pastori.
Protease problems
See the protease problems page.