- 专利标题: Enhanced protein expression and methods thereof
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申请号: US15771618申请日: 2016-10-27
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公开(公告)号: US11306319B2公开(公告)日: 2022-04-19
- 发明人: Cristina Bongiorni , Marguerite A. Cervin , George England , Chao Zhu , Frank Wouter Koopman
- 申请人: DANISCO US INC
- 申请人地址: US CA Palo Alto
- 专利权人: DANISCO US INC
- 当前专利权人: DANISCO US INC
- 当前专利权人地址: US CA Palo Alto
- 国际申请: PCT/US2016/059078 WO 20161027
- 国际公布: WO2017/075195 WO 20170504
- 主分类号: C12N15/00
- IPC分类号: C12N15/00 ; C12N15/75 ; C12N9/12 ; C12P21/02 ; C12P21/00 ; C12N1/20 ; C12R1/01
摘要:
The present disclosure is generally related to modified Gram positive bacterial cells producing increased amounts of one or more protein(s) of interest and modified Gram positive bacterial cells having increased genetic competency. Thus, certain embodiments of the disclosure are directed to modified Gram positive bacterial cells expressing an increased amount of a protein of interest, relative to an unmodified (parental) Gram positive bacterial cell expressing the same protein of interest, wherein the modified bacterial cell comprises at least one mutation in a rpoC gene encoding a variant RNA-polymerase (RNAP) β′-subunit polypeptide. In certain embodiments, the rpoC gene encoding the variant β′-subunit polypeptide is integrated into the chromosome of the modified cell. In other embodiments, the rpoC gene encoding the variant β′-subunit polypeptide is comprised on an extrachromosomal plasmid introduced into the modified cell. In other embodiments, the disclosure is directed to competent Bacillus host cells comprising at least one copy of a nucleic acid construct encoding a modified rpoC polypeptide comprising 90% sequence identity to SEQ ID NO: 8 and an aspartic acid to glycine substitution at position 796 of SEQ ID NO: 8, wherein the polynucleotide encoding the rpoC polypeptide is foreign to the Bacillus host cell that was non-competent prior to the introduction of the first nucleic acid construct.
公开/授权文献
- US20180327757A1 ENHANCED PROTEIN EXPRESSION AND METHODS THEREOF 公开/授权日:2018-11-15
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