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US 5,521,084 AGranted
Bovine heat shock promoter and uses thereof
AssigneeBiostar, Inc.
InventorsJacek Kowalski, Scott Gilbert, Timothy J. Zamb
Life of this patent
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Abstract
A novel expression system using the heat-inducible bovine hsp70A promoter and associated cis-acting elements is disclosed. The system provides for the continuous production of a highly pure, authentic protein, substantially free of infectious viral and cellular protein contaminants.
Claims19
A Madin-Darby bovine kidney (MDBK) cell transformed with a recombinant expression construct comprising:
- (a) a nucleic acid molecule that comprises a coding sequence for a selected polypeptide;
- and
- (b) hsp70 control sequences operably linked to said nucleic acid molecule, whereby said coding sequence will be transcribed and translated when in a host cell to produce said selected polypeptide, and at least one of said control sequences is heterologous to said coding sequence,
- wherein said hsp70 control sequences comprise
- (i) a bovine hsp70A promoter selected from the group consisting of a (1) a promoter which comprises the nucleotide sequence depicted at nucleotide positions 3.50 to 441, inclusive, of the upper strand of FIG. 2 (SEQ ID NO:1);
- (2) a promoter which comprises the nucleotide sequence depicted at nucleotide positions 265 to 441, inclusive, of the upper strand of FIG. 2 (SEQ ID NO:1) and (3) a promoter which comprises the nucleotide sequence depicted at nucleotide positions 1 to 441, inclusive, of the upper strand of FIG. 2 (SEQ ID NO:1),
- (ii) a bovine 5'-untranslated region positioned downstream of said promoter, and
- (iii) a 3'-untranslated region positioned downstream of said coding sequence,
5 more elements
A Madin-Darby bovine kidney (MDBK) cell transformed with a recombinant expression construct comprising:
- (a) a nucleic acid molecule that comprises a coding sequence for a selected polypeptide;
- and
- (b) hsp70 control sequences operably linked to said nucleic acid molecule and a 3'-untranslated region positioned downstream of said coding sequence, whereby said coding sequence will be transcribed and translated when in a host cell to produce said selected polypeptide, and at least one of said control sequences is heterologous to said coding sequence,
- wherein said hsp70 control sequences comprise the nucleotide sequence depicted at nucleotide positions 1 to 666, inclusive, of the upper strand of FIG. 2 (SEQ ID NO:1).
1 more element
Dependencies are inferred from the claim text.