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

Inventor： Xuqiu TAN ,  Corey DODGE ,  Kevin Li WONG ,  Aileen Aboy MILAN ,  Samantha ORCHARD ,  Ruorong CAI

IPC: C12Q1/6895 ,  C12N15/81 ,  C12P21/00

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.

公开(公告)号：US20210139871A1

公开(公告)日：2021-05-13

申请号：US16954276

申请日：2018-12-10

Applicant: BASF SE

Inventor： Xuqiu TAN ,  Kenneth BARRETT ,  Jared DENNIS ,  Dongmei REN ,  Corey DODGE ,  Bin ZHANG

IPC: C12N9/22 ,  C11D3/386 ,  A23L33/18 ,  A23K20/147 ,  A23L29/00 ,  A23K20/189

Abstract: Disclosed herein are polypeptides having nuclease activity. Some of the polypeptides having nuclease activity were generated by an improved gene site mutagenesis (“GSSM”) method or the tailored multi-site combinatorial assembly (“TMCA”) method. Also disclosed are compositions and kits comprising the polypeptides having nuclease activity, and methods for making and using these polypeptides, compositions and kits.