US4551433A
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Microbial hybrid promoters
Assignee: Genentech Inc
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Filing: 1982-01-11
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Granted: 1985-11-05
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Expires: 2002-01-11
Abstract
Abstract Novel microbial hybrid promoters and their use to direct the microbial expression of heterologous genes are described. Such promoters are selectively and functionally constructed by recombinant techniques, utilizing the discovery that certain DNA regions of given promoters are responsible for particularly advantageous functional properties.
Inventors
- 👤 Herman A. DeBoer
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-26 02:20
CPC Classifications
C; C12; C12P; C12P21/00; C12P21/02; C12N; C12N15/00; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/822; Y10S435/848; Y10S435/849
Claims Analysis
Total Claims: 18
Claim 1
Independent Claim
89 words
A hybrid promoter/operator capable of directing E.coli DNA-dependent RNA polymerase transcription of a recombinant DNA fragment positioned downstream therefrom comprising: (a) a portion of a promoter/operator sequence which portion is capable of being derepressed by induction and which promoter exhibits a first strength, and, (b) operably positioned upstream therefrom, a fragment comprising the -35 consensus sequence and 5' flanking region of a second promoter having a greater strength; elements (a) and (b) being covalently linked between about said -35 consensus sequence and the Prinbnow box of said promoter/operator sequence.
Claim 2
Dependent Claim
89 words
A hybrid promoter/operator capable of directing E.coli DNA-dependent RNA polymerase transcription of a recombinant DNA fragment positioned downstream therefrom comprising: (a) a portion of a promoter/operator sequence which portion is capable of being derepressed by induction and which promoter exhibits a first strength, and, (b) operably positioned upstream therefrom, a fragment comprising the -35 consensus sequence and 5' flanking region of a second promoter having a greater strength; elements (a) and (b) being covalently linked between about said -35 consensus sequence and the Prinbnow box of said promoter/operator sequence.
Claim 3
Dependent Claim
26 words
A hybrid promoter/operator according to claim 1 wherein said elements (a) and (b) are linked at a position between about nucleotide base pairs -10 to -35.
Claim 4
Dependent Claim
20 words
A hybrid promoter/operator according to claim 1 wherein said elements (a) and (b) are linked via complementary restriction sites thereon.
Claim 5
Dependent Claim
15 words
A hybrid promoter/operator according to claim 3 wherein said complementary restriction sites are synthetically derived.
Claim 6
Dependent Claim
37 words
A hybrid promoter/operator according to claims 1, 2, 3, or 4 wherein said element (a) is derived from a promoter operator selected from the group consisting of lac, gal P2, ara BAD, ara C and gal P1.
Claim 7
Dependent Claim
36 words
A hybrid promoter/operator according to claims 1, 2, 3, or 4 wherein said element (b) is derived from a promoter selected from the group consisting of trp, rrn, lambda PR, lambda PL, str, spc and rpoB.
Claim 8
Dependent Claim
29 words
An E. coli culture capable of producing a heterologous polypeptide by expression of a gene coding therefor, said expression being directed by a hybrid promoter/operator according to claim 1.
Claim 9
Dependent Claim
77 words
A hybrid promoter/operator which comprises: (a) an operator DNA sequence of a first promoter/operator which is capable of being derepressed by induction, and which comprises that sequence immediately downstream of a position between about -10 and -35 of said first promoter operator; and (b) upstream from the sequence of (a), a DNA sequence which comprises that sequence of a second promoter which is immediately upstream of a position between about -10 and -35 of said second promoter.
Claim 10
Dependent Claim
77 words
A hybrid promoter/operator which comprises: (a) an operator DNA sequence of a first promoter/operator which is capable of being derepressed by induction, and which comprises that sequence immediately downstream of a position between about -10 and -35 of said first promoter operator; and (b) upstream from the sequence of (a), a DNA sequence which comprises that sequence of a second promoter which is immediately upstream of a position between about -10 and -35 of said second promoter.
Claim 11
Dependent Claim
85 words
A hybrid promoter/operator capable of directing E.coli DNA-dependent RNA polymerase catalyzed transcription which comprises: (a) an upstream portion comprising the DNA sequence of a first promoter, which sequence is upstream of a position between about -10 and -35 of such first promoter; and (b) a downstream portion comprising the DNA sequence of a second promoter/operator which sequence is downstream of a position between about -10 and -35 of said second promter/operator, and which sequence contains an operator which is capable of being derepressed by induction.
Claim 12
Dependent Claim
85 words
A hybrid promoter/operator capable of directing E.coli DNA-dependent RNA polymerase catalyzed transcription which comprises: (a) an upstream portion comprising the DNA sequence of a first promoter, which sequence is upstream of a position between about -10 and -35 of such first promoter; and (b) a downstream portion comprising the DNA sequence of a second promoter/operator which sequence is downstream of a position between about -10 and -35 of said second promter/operator, and which sequence contains an operator which is capable of being derepressed by induction.
Claim 13
Dependent Claim
21 words
The hybrid promoter/operator of claims 1, 8, or 9 wherein the operator sequence is the operator sequence of the lac promoter/operator.
Claim 14
Dependent Claim
35 words
The hybrid promoter/operator of claim 10 which comprises the DNA sequence downstream from position -19 of the lac promoter/operator linked to, and downstream from, the DNA sequence upstream of position -22 of the trp promoter.
Claim 15
Dependent Claim
32 words
A hybrid promoter/operator according to claim 10 comprising a DNA sequence upstream from position -12 of the trp promoter linked to a DNA sequence which comprises the core of the lac operator.
Claim 16
Dependent Claim
41 words
The hybrid promoter/operator of claim 10 which comprises a DNA sequence downstream from position -19 of the lac promoter operator linked downstream from, and via a synthetic DNA sequence to, a DNA sequence upstream from position -28 of the rrn promoter.
Claim 17
Dependent Claim
27 words
A replicable expression vehicle comprising a heterologous gene insert, the expression of which is under the control of the hybrid promoter/operator of claims 1, 8, or 9.
Claim 18
Dependent Claim
11 words
An E.coli strain transformed with the vehicle according to claim 14.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1988-11-04 | 1989-05-06 | 1989-11-05 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1992-11-04 | 1993-05-06 | 1993-11-05 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1996-11-04 | 1997-05-06 | 1997-11-05 | $7700.00 | Window Closed / Lapsed |