Yeast expression purification
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Last Update: 2021-01-24
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Source: Internet
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Author: User
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asked:
yeast
after the methanol time period induction, and then centrifugation, the resulting upper clear is protein? Do you need a NI post? Need to break? Thank you for your guidance, I do not know the expression and purification of yeast, only done the expression and purification of the true core and the original, now I do not know whether the purification of yeast and the purification of the general core (soluble expression products) is not the same, thank you, I enzyme Mother-positive recombination son has been screened out, the next step has to do induction, but the yeast expression purification is not at all, annoying ah
: I don't know what you mean by positive clones that have been screened out. Yes
PCR
identified your purpose
gene
, or is it just a clone screened on an RDB or MD or G418 tablet? If this is done, you can refer to the methods in Invitrogen's Bich Yeast Expression Manual for induced expression. Directly from the tablet to pick more clones, with MGY or BMGY
culture
-based culture, and then with BMMY
culture
induced expression,
SDS
-PAGE to detect whether the purpose protein is expressed, while selecting high-expression strains, while using Western-blotting to identify whether your purpose protein is ok.
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