Engineered ribosomal promoters for protein production in microorganisms
AssigneeDanisco US Inc.
InventorsCristina Bongiorni, Marc Kolkman, Chris Leeflang, Virgil Arthur Rhodius, Anita Van Kimmenade
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Abstract
The instant disclosure is generally related to novel engineered (hybrid) promoters. In certain embodiments, the instant disclosure is directed to one or more nucleic acid compositions comprising engineered promoters operably linked to nucleic acids encoding proteins of interest. Thus, the disclosure set forth herein described methods and compositions for the production of proteins of interest using one or more novel engineered (hybrid) promoters of the disclosure.
Claims14
An isolated nucleic acid comprising an engineered hybrid promoter operably linked to a nucleic acid encoding a protein of interest (POI), wherein the hybrid promoter comprises a nucleotide sequence having at least 98% identity to any one of SEQ ID NO:
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A method for screening transformed host cells for increased expression of a POI comprising:
- (i) transforming a host cell with an isolated nucleic acid comprising a heterologous engineered hybrid promoter operably linked to a nucleic acid encoding a protein of interest (POI), wherein the hybrid promoter comprises the nucleotide sequence of any one of SEQ ID NO:
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- (ii) transforming a host cell with an isolated nucleic acid comprising its native promoter operably linked to a nucleic acid encoding the same POI as step (i), wherein the host cells transformed in steps (i) and (ii) are host cells of the same Genus species and genetic background, and
- (iii) culturing the modified cells under conditions such that the POI is expressed, wherein an increase in the expression of the POI coding sequence in step (i), relative to the expression of the same POI coding sequence in step (ii), indicates increased expression of the POI.
A method for increasing the expression of a POI in a host cell comprising:
- (i) modifying a host cell by introducing into the host cell a nucleic acid comprising an engineered hybrid promoter operably linked to a nucleic acid encoding a protein of interest (POI), wherein the hybrid promoter comprises the nucleotide sequence of any one of SEQ ID NO:
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- (ii) culturing the modified host cell under conditions such that the POI is expressed.
Dependencies are inferred from the claim text.