US4503155A
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Multifunctional, cloning vectors for use in Streptomyces, Bacillus, and E. coli
Assignee: Eli Lilly and Co
•
Filing: 1982-02-01
•
Granted: 1985-03-05
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Expires: 2002-02-01
Abstract
Abstract The present invention discloses multifunctional recombinant DNA cloning vectors for use in Streptomyces, Bacillus, and E. coli. The invention further discloses transformants of the aforementioned vectors.
Inventors
- 👤 James R. Miller
- 👤 Jeffrey T. Fayerman
- 👤 Steven Kovacevic
- 👤 Nancy E. Beerman
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-24 14:29
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/74; C12N15/75; C12N15/76; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/822; Y10S435/832; Y10S435/848; Y10S435/849; Y10S435/886
Claims Analysis
Total Claims: 54
Claim 1
Independent Claim
127 words
A multifunctional recombinant DNA cloning vector comprising: (a) two or more functionally different origins of replication that are independently selected from the group consisting of any origin of replication that is functional in Streptomyces, any origin of replication that is functional in Bacillus, and any origin of replication that is functional in E. coli, and . (b) one or more DNA segments that confer resistance to at least one antibiotic when transformed into a sensitive host cell in which an origin of replication comprising said vector is functional, said host cells being susceptible to transformation, cell division, and culture, subject to the limitatidn that when said vector is limited to two functionally different origins of replication, neither of the origins of replication is functional in E. coli.
Claim 2
Dependent Claim
127 words
A multifunctional recombinant DNA cloning vector comprising: (a) two or more functionally different origins of replication that are independently selected from the group consisting of any origin of replication that is functional in Streptomyces, any origin of replication that is functional in Bacillus, and any origin of replication that is functional in E. coli, and . (b) one or more DNA segments that confer resistance to at least one antibiotic when transformed into a sensitive host cell in which an origin of replication comprising said vector is functional, said host cells being susceptible to transformation, cell division, and culture, subject to the limitatidn that when said vector is limited to two functionally different origins of replication, neither of the origins of replication is functional in E. coli.
Claim 3
Dependent Claim
43 words
The cloning vector of claim 1 wherein the origins of replication that are functional in Streptomyces, Bacillus, and E. coli are respectively a restriction fragment of plasmid pEL103, a restriction fragment of plasmid pHI-16, and a restriction fragment of an E. coli plasmid.
Claim 4
Dependent Claim
30 words
The cloning vector of claim 1 wherein the origins of replication in said vectors are limited to a restriction fragment of plasmid pEL103 and a restriction fragment of plasmid pHI-16.
Claim 5
Dependent Claim
17 words
The cloning vector of claim 1, 2 or 3 wherein one DNA segment conveys resistance to thiostrepton.
Claim 6
Dependent Claim
17 words
The cloning vector of claim 1, 2 or 3 wherein one DNA segment conveys resistance to neomycin.
Claim 7
Dependent Claim
17 words
The cloning vector of claim 1, 2 or 3 wherein one DNA segment conveys resistance to chloramphenicol.
Claim 8
Dependent Claim
17 words
The cloning vector of claim 1, 2 or 3 wherein one DNA segment conveys resistance to kanamycin.
Claim 9
Dependent Claim
14 words
The cloning vector of claim 2 wherein one DNA segment conveys resistance to ampicillin.
Claim 10
Dependent Claim
20 words
The cloning vector of claim 4 wherein the DNA segment is the ˜1.6 kb BamHI restriction fragment of plasmid pLR2.
Claim 11
Dependent Claim
20 words
The cloning vector of claim 5 wherein the DNA segment is the ˜3.4 kb BamHI restriction fragment of plasmid pLR1.
Claim 12
Dependent Claim
20 words
The cloning vedtor of claim 6 wherein the DNA segment is the ˜4.6 kb BamHI restriction fragment of plasmid pHI-16.
Claim 13
Dependent Claim
20 words
The cloning vector of claim 7 wherein the DNA segment is the ˜3.9 kb BamHI restriction fragment of plasmid pHI-18.
Claim 14
Dependent Claim
39 words
The cloning vector of claim 1, 2 or 3 wherein the one or more DNA segments that convey antibiotic resistance are selected from the group consisting of segments that convey resistance to hygromycin, viamycin, tylosin, erythromycin, streptomycin, and bacteriocin.
Claim 15
Dependent Claim
23 words
The cloning vector of claim 3 wherein the origin of replication containing restriction fragment of plasmid pEL103 is the ˜2.8 kb BamHI fragment.
Claim 16
Dependent Claim
24 words
The cloning vector of claim 2 in which the E. coli plasmid is selected from the group of plasmids consisting of pBR322 and pBR328.
Claim 17
Dependent Claim
77 words
The cloning vector of claim 1 which is selected from the group of plasmids consisting of PBS1, pBS2, pBS3, pBS4, pBS5, pBS6, pBS7, pBS8, pBS9, pBS10, pBS11, pBS12, pBS13, pBS14, pBS15, pBS16, pBS17, pBS18, pBS19, pBS20, pBS21, pBS22, pBS23, pBS24, pBS25, pBS26, pBS27, pBS28, pBS29, pBS30, pBS31, pBS32, pBS33, pBS34, pBS35, pBS36, pBS37, pBS38, pBS39, pBS40, pBS41, pBS42, pBS43, pBS44, pBS45, pBS46, pBS47, pBS48, pBS49, pBS50, pBS51, pBS52, pBS53, pBS54, pBS55, pBS56, pBS57, pBS58, pBS59, and pBS60.
Claim 18
Dependent Claim
10 words
The cloning vector of claim 16 which is plasmid pBS1.
Claim 19
Dependent Claim
9 words
The cloning vedtor of claim 16 which is pBS2.
Claim 20
Dependent Claim
9 words
The cloning vector of claim 16 which is pBS3.
Claim 21
Dependent Claim
9 words
The cloning vector of claim 16 which is pBS4.
Claim 22
Dependent Claim
9 words
The cloning vector of claim 16 which is pBS5.
Claim 23
Dependent Claim
9 words
The cloning vector of claim 16 which is pBS7.
Claim 24
Dependent Claim
9 words
The cloning vector of claim 16 which is pBS8.
Claim 25
Dependent Claim
9 words
The cloning vector of claim 16 which is pBS9.
Claim 26
Dependent Claim
9 words
The cloning vector of claim 16 which is pBS10.
Claim 27
Dependent Claim
9 words
The cloning vector of claim 16 which is pBS49.
Claim 28
Dependent Claim
9 words
The cloning vector of claim 16 which is pBS60.
Claim 29
Dependent Claim
60 words
A transformed restrictionless host cell comprising a recombinant DNA cloning vector of claim 1 subject to the limitation that said host cell, prior to transformation, is sensitive to an antibiotic for which resistance is conferred by a DNA segment comprising said vector, and further that at least one origin of replication comprising said vector is functional in said host cell.
Claim 30
Dependent Claim
85 words
The transformed restrictionless host cell of claim 28 in which the recombinant DNA cloning vector is selected from the group of plasmids consisting of pBS1, pBS2, pBS3, pBS4, pBS5, pBS6, pBS7, pBS8, pBS9, pBS10, pBS11, pBS12, pBS13, pBS14, pBS15, pBS16, pBS17, pBS18, pBS19, pBS20, pBS21, pBS22, pBS23, pBS24, pBS25, pBS26, pBS27, pBS28, pBS29, pBS30, pBS31, pBS32, pBS33, pBS34, pBS35, pBS36, pBS37, pBS38, pBS39, pBS40, pBS41, pBS42, pBS43, pBS44, pBS45, pBS46, pBS47, pBS48, pBS49, pBS50, pBS51, pBS52, pBS53, pBS54, pBS55, pBS56, pBS57, pBS58, pBS59, and pBS60.
Claim 31
Dependent Claim
11 words
The transformed host cell of claim 28 which is a Streptosporangium.
Claim 32
Dependent Claim
11 words
The transformed host cell of claim 28 which is a Actinoplanes.
Claim 33
Dependent Claim
11 words
The transformed host cell of claim 28 which is a Nocardia.
Claim 34
Dependent Claim
11 words
The transformed host cell of claim 28 which is a Micromonospora.
Claim 35
Dependent Claim
11 words
The transformed host cell of claim 28 which is a Staphylococcus.
Claim 36
Dependent Claim
13 words
The transformed host cell of claim 28 or 29 which is a Streptomyces.
Claim 37
Dependent Claim
13 words
The transformed host cell of claim 28 or 29 which is a Bacillus.
Claim 38
Dependent Claim
13 words
The transformed host cell of claim 28 or 29 which is E. coli.
Claim 39
Dependent Claim
11 words
The transformed host cell of claim 29 which is Streptomyces fradiae.
Claim 40
Dependent Claim
11 words
The transformed host cell of claim 29 which is Streptomyces coelicolor.
Claim 41
Dependent Claim
11 words
The transformed host cell of claim 29 which is Streptomyces ambofaciens/pBS1.
Claim 42
Dependent Claim
11 words
The transformed host cell of claim 29 which is Streptomyces ambofaciens/pBS2.
Claim 43
Dependent Claim
11 words
The transformed host cell of claim 29 which is Streptomyces ambofaciens/pBS5.
Claim 44
Dependent Claim
11 words
The transformed host cell of claim 29 which is Streptomyces ambofaciens/pBS7.
Claim 45
Dependent Claim
12 words
The transformed host cell of claim 29 which is Bacillus subtilis MI112/pBS1.
Claim 46
Dependent Claim
12 words
The transformed host cell of claim 29 which is Bacillus subtilis MI112/pBS2.
Claim 47
Dependent Claim
12 words
The transformed host cell of claim 29 which is Bacillus subtilis MI112/pBS5.
Claim 48
Dependent Claim
12 words
The transformed host cell of claim 29 which is Bacillus subtilis MI112/pBS7.
Claim 49
Dependent Claim
11 words
The transformed host cell of claim 29 E. coli K12 HB101/pBS9.
Claim 50
Dependent Claim
11 words
The transformed host cell of claim 29 which is Streptomyces ambofaciens/pBS9.
Claim 51
Dependent Claim
12 words
The transformed host cell of claim 29 which is Bacillus subtilis MI112/pBS9.
Claim 52
Dependent Claim
16 words
A recombinant DNA cloning vector selected from the group of plasmids consisting of pHI-16 and pHI-18.
Claim 53
Dependent Claim
16 words
A recombinant DNA cloning vector selected from the group of plasmids consisting of pHI-16 and pHI-18.
Claim 54
Dependent Claim
18 words
A Bacillus subtilis MI112 host cell that is transformed with a recombinant DNA cloning vector of claim 51.
References Cited (Backward Citations)
US4340674A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1988-03-04 | 1988-09-03 | 1989-03-05 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1992-03-04 | 1992-09-03 | 1993-03-05 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1996-03-04 | 1996-09-03 | 1997-03-05 | $7700.00 | Window Closed / Lapsed |