US4921801A
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Novel recombinant DNA cosmid shuttle vectors
Assignee: Eli Lilly and Co
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Filing: 1986-03-20
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Granted: 1990-05-01
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Expires: 2006-03-20
Abstract
Abstract Novel recombinant DNA cosmid shuttle vectors and a method of using them in the construction of genomic DNA libraries are described. The vectors demonstrate the incorporation of both the size selection and in vitro packaging mechanisms of lambda into a Streptomyces-E. coli shuttle vector by the incorporation of two or more COS sequences of bacteriophage lambda.
Inventors
- 👤 R. Nagaraja Rao
- 👤 Richard K. Stanzak
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-09 00:07
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/74; C12N15/76; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/822; Y10S435/848; Y10S435/849; Y10S435/886; Y10S435/889
Claims Analysis
Total Claims: 59
Claim 1
Independent Claim
65 words
A recombinant double standard DNA cosmid shuttle vector comprising: (a) a replicon that is functional in E. coli, (b) a replicon that is functional in Streptomyces, (c) a double stranded DNA segment that contains two or more cos sequences of bacteriophage lambda, and (d) one or more DNA segments that convey resistance to at least one antibiotic when transformed into a sensitive restrictionless host cell.
Claim 2
Dependent Claim
65 words
A recombinant double standard DNA cosmid shuttle vector comprising: (a) a replicon that is functional in E. coli, (b) a replicon that is functional in Streptomyces, (c) a double stranded DNA segment that contains two or more cos sequences of bacteriophage lambda, and (d) one or more DNA segments that convey resistance to at least one antibiotic when transformed into a sensitive restrictionless host cell.
Claim 3
Dependent Claim
22 words
The recombinant DNA cosmid shuttle vector of claim 1 selected from the group consisting of pKC420, pKC427, pKC428, pKC448, pKC462, and pKC467.
Claim 4
Dependent Claim
12 words
The recombinant DNA cosmid shuttle vector of claim 2 which is pKC420.
Claim 5
Dependent Claim
12 words
The recombinant DNA cosmid shuttle vector of claim 2 which is pKC427.
Claim 6
Dependent Claim
12 words
The recombinant DNA cosmid shuttle vector of claim 2 which is pKC428.
Claim 7
Dependent Claim
12 words
The recombinant DNA cosmid shuttle vector of claim 2 which is pKC448.
Claim 8
Dependent Claim
12 words
The recombinant DNA cosmid shuttle vector of claim 2 which is pKC462.
Claim 9
Dependent Claim
12 words
The recombinant DNA cosmid shuttle vector of claim 2 which is pKC467.
Claim 10
Dependent Claim
30 words
The recombinant DNA cosmid shuttle vector of claim 1 wherein the E. coli replicon is selected from the group consisting of replicon-containing fragments of plasmids pBR322, pBR324, pBR325 and pBR328.
Claim 11
Dependent Claim
30 words
The recombinant DNA cosmid shuttle vector of claim 1 wherein the Streptomyces replicon is selected from the group consisting of replicon-containing fragments of plasmids SCP2, SCP2*, SLP1, pEL103 and pFJ265.
Claim 12
Dependent Claim
28 words
A transformed host cell selected from the group consisting of restrictionless Streptomyces and E. coli, said host cell comprising a recombinant DNA cosmid shuttle vector of claim 1.
Claim 13
Dependent Claim
28 words
A transformed host cell selected from the group consisting of restrictionless Streptomyces and E. coli, said host cell comprising a recombinant DNA cosmid shuttle vector of claim 2.
Claim 14
Dependent Claim
11 words
The transformed host cell of claim 12 which is restrictionless Streptomyces.
Claim 15
Dependent Claim
30 words
The transformed host cell of claim 13 which is selected from the group consisting of Streptomyces ambofaciens, Streptomyces aureofaciens, Streptomyces cinnamonesis, Streptomyces fradiae, Streptomyces granuloruber, Streptomyces lividans, and Streptomyces griseofuscus.
Claim 16
Dependent Claim
11 words
The transformed host cell of claim 13 which is Streptomyces ambofaciens/pKC420.
Claim 17
Dependent Claim
11 words
The transformed host cell of claim 13 which is Streptomyces ambofaciens/pKC448.
Claim 18
Dependent Claim
11 words
The transformed host cell of claim 13 which is Streptomyces ambofaciens/pKC462.
Claim 19
Dependent Claim
11 words
The transformed host cell of claim 13 which is Streptomyces fradiae/pKC420.
Claim 20
Dependent Claim
11 words
The transformed host cell of claim 13 which is Streptomyces fradiae/pKC448.
Claim 21
Dependent Claim
11 words
The transformed host cell of claim 13 which is Streptomyces lividans/pKC420.
Claim 22
Dependent Claim
11 words
The transformed host cell of claim 13 which is Streptomyces lividans/pKC448.
Claim 23
Dependent Claim
11 words
The transformed host cell of claim 13 which is Streptomyces lividans/pKC462.
Claim 24
Dependent Claim
11 words
The transformed host cell of claim 13 which is Streptomyces lividans/pKC467.
Claim 25
Dependent Claim
11 words
The transformed host cell of claim 12 which is E. coli.
Claim 26
Dependent Claim
13 words
The transformed host cell of claim 24 which is E. coli K12 SF8/pKC420.
Claim 27
Dependent Claim
13 words
The transformed host cell of claim 24 which is E. coli K12 SF8/pKC448.
Claim 28
Dependent Claim
12 words
The transforled host cell of claim 24 which is E. coli DH1/pKC462.
Claim 29
Dependent Claim
58 words
A method for using the cosmid shuttle vector of claim 1 to construct genomic DNA libraries, said method comprising: (a) ligating a genomic DNA segment into said cosmid shuttle vector, (b) packaging said ligated cosmid into bacteriophage lambda particles, (c) transducing said packaged cosmid into E. coli, and (d) transforming the recombinant cosmid into a Streptomyces host cell.
Claim 30
Dependent Claim
26 words
A method according to claim 28, wherein said cosmic shuttle vector DNA is selected from the group consisting of pKC420 pKC427, pKC428, pKC448, pKC462 and pKC467.
Claim 31
Dependent Claim
12 words
The method of claim 29, wherein said cosmid shuttle vector is p-KC420.
Claim 32
Dependent Claim
12 words
The method of claim 29, wherein said cosmid shuttle vector is p-KC448.
Claim 33
Dependent Claim
12 words
The method of claim 29 wherein said cosmid shuttle vector is pKC-462.
Claim 34
Dependent Claim
12 words
The method of claim 29 wherein said cosmid shuttle vector is pKC467.
Claim 35
Dependent Claim
12 words
The recombinant DNA cosmid shuttle vector of claim 1 which is pKC462A.
Claim 36
Dependent Claim
12 words
The recombinant DNA cosmid shuttle vector of claim 1 which is pKC467A.
Claim 37
Dependent Claim
12 words
The recombinant DNA cosmid shuttle vector of claim 1 which is cos111.
Claim 38
Dependent Claim
11 words
The transformed host cell of claim 11 which is Streptomyces ambofaciens/pKC462A.
Claim 39
Dependent Claim
11 words
The transformed host cell of claim 11 which is Streptomyces lividans/cos111.
Claim 40
Dependent Claim
13 words
The transformed host cell of claim 11 which is E. coli K12 SF8/pKC462A.
Claim 41
Dependent Claim
13 words
The transformed host cell of claim 11 which is E. coli K12 SF8/pKC467A.
Claim 42
Dependent Claim
13 words
The transformed host cell of claim 11 which is E. coli K12 SF8/cos111.
Claim 43
Dependent Claim
20 words
The method of claim 28 wherein said cosmid shuttle vector is selected from the group consisting of pKC462A and cos111.
Claim 44
Dependent Claim
22 words
The recombinant DNA cosmid shuttle vector of claim 1 which is selected from the group consisting of pKC513, pKC531, pKC532, and pKC578.
Claim 45
Dependent Claim
12 words
The recombinant DNA cosmid shuttle vector of claim 43 which is pKC513.
Claim 46
Dependent Claim
12 words
The recombinant DNA cosmid shuttle vector of claim 43 which is pKC531.
Claim 47
Dependent Claim
12 words
The recombinant DNA cosmid shuttle vector of claim 43 which is pKC532.
Claim 48
Dependent Claim
12 words
The recombinant DNA cosmid shuttle vector of claim 43 which is pKC578.
Claim 49
Dependent Claim
10 words
The transformed host of claim 11 that is Streptomyces griseofuscus.
Claim 50
Dependent Claim
10 words
The transformed host of claim 48 that is Streptomyces griseofuscus/pKC531.
Claim 51
Dependent Claim
10 words
The transformed host of claim 48 that is Streptomyces griseofuscus/pKC532.
Claim 52
Dependent Claim
10 words
The transformed host of claim 48 that is Streptomyces griseofuscus/pKC578.
Claim 53
Dependent Claim
21 words
The recombinant DNA cosmid shuttle vector of claim 1 which further comprises a DNA segment that contains a Strepomyces fertility factor.
Claim 54
Dependent Claim
17 words
The recombinant DNA cosmid shuttle vector of claim 52 wherein the fertility factor is from plasmid SCP2*.
Claim 55
Dependent Claim
12 words
The recombinant DNA cosmid shuttle vector of claim 53 which is pKC505.
Claim 56
Dependent Claim
28 words
A transformed host cell selected from the group consisting of restrictionless Streptomcyes and E. coli, said host cell comprising the recombinant DNA cosmid shuttle vector of claim 54.
Claim 57
Dependent Claim
11 words
The transformed host cell of claim 55 which is Streptomyces griseofuscus/pKC505.
Claim 58
Dependent Claim
13 words
The transformed host cell of claim 55 which is E. coli K12 DH1/pKC505.
Claim 59
Dependent Claim
21 words
The method of claim 28 wherein said cosmid shuttle vector is selected from the group consisting of pKC505, pKC531, and pKC578.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1993-04-30 | 1993-10-30 | 1994-05-01 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1997-04-30 | 1997-10-30 | 1998-05-01 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2001-04-30 | 2001-10-30 | 2002-05-01 | $7700.00 | Window Closed / Lapsed |