DNA construct for regulating the expression of a polypeptide coding sequence in a transformed bacterial host cell
AssigneeMonsanto Technology LLC
InventorsGregg Bogosian, Julia P. O'Neil, Katherine C. Terlesky
Life of this patent
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Abstract
A heterologous polypeptide is expressed under the control of a DNA construct containing a promoter region derived from a cyanophage or cyanobacteria promoter. In one embodiment, such a promoter region is operably linked to an operator region derived from an operator native to the host cell. In another embodiment, the operator region is positioned upstream of the promoter region.
Claims26
A DNA construct for regulating the expression of a heterologous polypeptide coding sequence in a transformed bacterial host cell, comprising:
- (a) a heterologous cyanophage N-1 early gene promoter, operably linked to
- (b) an operator segment from an operator native to the host cell, wherein the operator segment is located upstream of the promoter region.
A DNA construct for regulating the expression of a heterologous polypeptide coding sequence in a transformed bacterial host cell, comprising
- a heterologous promoter, operably linked to an operator segment from an operator native to the host cell, wherein the operator segment is positioned upstream of the promoter;
- wherein said operator is capable of binding to a repressor protein made by the host cell;
- and,
- wherein the promoter is capable of binding to an RNA polymerase produced by the bacterial host to initiate transcription, and
- wherein the promoter is a cyanophage N-1 early gene or Rhodobacter capsulatus promoter.
An expression vector comprising a DNA construct for regulating the expression of a polypeptide coding sequence in a transformed bacterial host cell, comprising:
- (a) a heterologous cyanophage N-1 early gene promoter, operably linked to
- (b) an operator segment from an operator native to the host cell, wherein the operator segment is located upstream of the promoter region.
A bacteria transformed with a heterologous polypeptide coding sequence, wherein the bacteria further comprises a DNA construct for regulating the expression of the heterologous polypeptide coding sequence, wherein the DNA construct comprises:
- (a) a heterologous cyanophage N-1 early gene promoter, operably linked to
- (b) an operator segment from an operator native to the bacteria, wherein the operator segment is located upstream of the promoter.
A method for producing a heterologous polypeptide in a transformed bacterial host comprising:
- (a) stably transforming a bacterial host with a vector containing a heterologous polypeptide coding sequence operably linked to a DNA construct for regulating the expression of the coding sequence, wherein the DNA construct comprises:
- (i) a heterologous cyanophage N-1 early gene promoter, operably linked to
- (ii) an operator segment from an operator native to the host cell, wherein the operator segment is located upstream of the promoter;
- (b) culturing the transformed bacteria under conditions that induce the expression of the coding sequence.
Dependencies are inferred from the claim text.