Methods for Identifying Promoters for Protein Production in Yeast

    公开(公告)号:US20210230708A1

    公开(公告)日:2021-07-29

    申请号:US15734768

    申请日:2019-06-03

    Applicant: BASF SE

    Abstract: Disclosed herein are methods of identifying promoters that drive protein expression independently of methanol and are useful in driving protein expression in yeast. The method may comprise the steps of: fermenting yeast cells under at least one fermentation condition in the absence of methanol, collecting samples at different times during fermentation under the at least one fermentation conditions, determining the relative mRNA levels associated with native yeast genes in the samples, identifying one or more of the native yeast genes associated with higher than average levels of mRNA and determining putative promoters associated with the higher than average levels of mRNA encoding the native yeast genes, making expression constructs, each construct comprising one of the identified putative promoters and a gene encoding a marker protein and introducing the expression constructs into yeast cells, culturing the yeast cells comprising the expression constructs in the absence of methanol, determining marker protein expression by the cultured yeast cells and comparing marker protein expression driven by each of the putative promoters to identify promoters that drive protein expression independently of methanol and are useful in driving protein expression in yeast.

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