Bovine heat shock promoter and uses thereof
AssigneeBiostar, Inc.
InventorsJacek Kowalski, Scott Gilbert, Timothy J. Zamb
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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.
Claims39
An isolated bovine hsp70A promoter capable of directing the transcription of a heterologous coding sequence positioned downstream therefrom, wherein said promoter is selected from the group consisting of (a) a promoter comprising the nucleotide sequence depicted at nucleotide positions 350 to 441, inclusive, of the upper strand of FIG. 2 (SEQ ID NO:1);
- (b) a promoter comprising the nucleotide sequence depicted at nucleotide positions 265 to 441, inclusive, of the upper strand of FIG. 2 (SEQ ID NO:1);
- (c) a promoter comprising the nucleotide sequence depicted at nucleotide positions 1 to 441, inclusive, of the upper strand of FIG. 2 (SEQ ID NO:1);
- and (d) a promoter comprising a nucleotide sequence that hybridizes to the promoter of (a), (b) or (c) in a Southern hybridization reaction performed under stringent conditions.
An isolated bovine hsp70A 5'-untranslated region.
A recombinant expression construct effective in directing the transcription of a selected coding sequence, said expression construct comprising:
- (a) bovine hsp70A control sequences comprising the nucleotide sequence depicted at nucleotide positions 1 to 666, inclusive, of the upper strand of FIG. 2 (SEQ ID NO:1), or a nucleotide sequence that hybridizes thereto in a Southern hybridization reaction performed under stringent conditions;
- and
- (b) a coding sequence operably linked to said control sequences, whereby said coding sequence can be transcribed and translated in a host cell, and at least one of said control sequences is heterologous to said coding sequence.
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).
Dependencies are inferred from the claim text.