Control of gene expression in eukaryotes
First Claim
Patent Images
1. A system for modulating expression of a target nucleic acid sequence comprising:
- (a) a first nucleic acid sequence operably linked to and regulates the expression of a second nucleic acid sequence, said second nucleic acid sequence encoding a regulator polypeptide; and
(b) a third nucleic acid sequence operably linked to and regulates the expression of a target nucleic acid sequence, whereby said regulator polypeptide binds to said third nucleic acid sequence and modulates the expression of said target nucleic acid sequence, and wherein said regulator polypeptide comprises (i) the amino acid sequence of SEQ ID NO;
2;
or (ii) an amino acid sequence encoded by a nucleotide sequence that hybridizes to the complement of the nucleotide sequence from 295 to 1035 of SEQ ID NO;
1 under hybridization conditions comprising incubating at 37°
C. in 1×
SSC and 50% formamide, and/or, wherein said third nucleic acid sequence (i) comprises the nucleotide sequence from 1225 to 1260 of SEQ ID NO;
1;
or (ii) hybridizes to the complement of the nucleotide sequence from 1225 to 1260 of SEQ ID NO;
1 under hybridizing conditions comprising incubating at 37°
C. in 1×
SSC and 50% formamide.
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Abstract
A chemically inducible gene expression system is described. A chimeric gene having a first sequence comprising a promoter and a regulator polypeptide is linked with a second sequence comprising a promoter and a coding or non-coding sequence. Expression of the target gene of the second sequence is controlled by the regulator polypeptide which is acted upon by an inducer. The inducer is a chemical compound, such as OHP, which acts upon the OHP responsive regulator polypeptide, which can be obtained from Rhodococcus sp. V49. Various domain regions and complementary response elements are also described.
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Citations
35 Claims
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1. A system for modulating expression of a target nucleic acid sequence comprising:
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(a) a first nucleic acid sequence operably linked to and regulates the expression of a second nucleic acid sequence, said second nucleic acid sequence encoding a regulator polypeptide; and
(b) a third nucleic acid sequence operably linked to and regulates the expression of a target nucleic acid sequence, whereby said regulator polypeptide binds to said third nucleic acid sequence and modulates the expression of said target nucleic acid sequence, and wherein said regulator polypeptide comprises (i) the amino acid sequence of SEQ ID NO;
2;
or (ii) an amino acid sequence encoded by a nucleotide sequence that hybridizes to the complement of the nucleotide sequence from 295 to 1035 of SEQ ID NO;
1 under hybridization conditions comprising incubating at 37°
C. in 1×
SSC and 50% formamide, and/or, wherein said third nucleic acid sequence (i) comprises the nucleotide sequence from 1225 to 1260 of SEQ ID NO;
1;
or (ii) hybridizes to the complement of the nucleotide sequence from 1225 to 1260 of SEQ ID NO;
1 under hybridizing conditions comprising incubating at 37°
C. in 1×
SSC and 50% formamide.- View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 32, 33, 34, 35)
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31. A plasmid deposited under NCIMB Accession No. 40997.
Specification